Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 194
Filter
1.
Article in Chinese | WPRIM | ID: wpr-880137

ABSTRACT

OBJECTIVE@#To investigate the value of CD44@*METHODS@#Flow cytometry was used to detected the proportion of CD44@*RESULTS@#The percentage of CD44@*CONCLUSION@#HCD44


Subject(s)
Flow Cytometry , Humans , Hyaluronan Receptors , Induction Chemotherapy , Leukemia, Myeloid, Acute/drug therapy , Neoplasm, Residual , Prognosis , Spleen
2.
Braz. oral res. (Online) ; 35: e073, 2021. tab, graf
Article in English | LILACS, BBO | ID: biblio-1278595

ABSTRACT

Abstract The aim of this study was to identify tumor parenchyma cells exhibiting immunohistochemical profile of stem cells by evaluating the immunoreactivity of OCT4 and CD44 in a number of cases of salivary gland neoplasms. The sample consisted of 20 pleomorphic adenomas, 20 mucoepidermoid carcinomas, and 20 adenoid cystic carcinomas located in major and minor salivary glands. The expression of OCT4 and CD44 was evaluated by the percentage of positive cells and the intensity of expression. All studied cases showed positive expression of OCT4 and CD44 and higher values than the control groups. For OCT4, luminal and non-luminal cells were immunostained in the case of pleomorphic adenomas and adenoid cystic carcinomas. Moreover, the immunoreactivity of CD44 was particularly evident in the non-luminal cells of these lesions. In mucoepidermoid carcinomas, there was immunoreactivity for both markers in squamous and intermediate cells and absence of staining in mucous cells. For both markers, a significantly higher immunostaining was verified in neoplasms located in the major salivary glands compared with lesions in minor salivary glands (p<0.001). In the total sample and in minor salivary glands, malignant neoplasms exhibited higher immunoreactivity for OCT4 than pleomorphic adenoma. A significant moderate positive correlation (r = 0.444 and p ≤ 0.001) was found between OCT4 and CD44 immunoexpression in the total sample. The high expression of OCT4 and CD44 may indicate that these proteins play an important role in identifying tumor stem cells.


Subject(s)
Humans , Salivary Gland Neoplasms , Carcinoma, Mucoepidermoid , Carcinoma, Adenoid Cystic , Adenoma, Pleomorphic , Hyaluronan Receptors/genetics , Octamer Transcription Factor-3/economics , Immunohistochemistry
3.
J. oral res. (Impresa) ; 9(6): 449-456, dic. 31, 2020. ilus, tab
Article in English | LILACS | ID: biblio-1178938

ABSTRACT

Objetive: To determine the expressions of the bone surface marker CD44 in samples of alveolar bone previously regenerated with allograft, xenograft, and mixed, using the technique of guided bone regeneration. Material and Methods: This exploratory study was approved by the institutional research and ethics committee. By means of intentional sampling and after obtaining informed consent for tissue donation, 20 samples of alveolar bone previously regenerated with guided bone regeneration therapy with particulate bone graft and membrane were taken during implant placement. The samples were stained with hematoxylin-eosin for histological analysis, and by immunohistochemistry for the detection of CD44. Results: Sections with hematoxylin-eosin showed bone tissue with the presence of osteoid matrix and mature bone matrix of usual appearance. Of the CD44+ samples, 80% were allograft and 20% xenograft. The samples with allograft-xenograft were negative. There were no differences in the intensity of CD44 expression between the positive samples. The marker was expressed in osteocytes, stromal cells, mononuclear infiltrate, and some histiocytes. Eighty percent of the CD44+ samples and 100% of the samples in which 60 or more cells were labelled corresponded to allografts (p=0.000). A total of 67% of the samples from the anterior sector, and 40% from the posterior sector were CD44+ (p=0.689). Conclusion: This study shows for the first time that guided bone regeneration using allografts is more efficient for the generation of mature bone determined by the expression of CD44, compared to the use of xenografts and mixed allograft-xenograft, regardless of the regenerated anatomical area.


Objetivo: Determinar la expresión del marcador de membrana óseo CD44 en muestras de hueso alveolar previamente regenerado con aloinjerto, xenoinjerto y mezcla mediante la técnica de regeneración ósea guiada. Material y Métodos: Con aval del Comité de Investigación y Ética, se realizó un estudio exploratorio. Por muestreo intencional y firma de consentimiento informado de donación, se tomaron durante la colocación del implante, 20 muestras de hueso alveolar previamente regenerado con terapia de regeneración ósea guiada con injerto óseo particulado y membrana. Las muestras fueron teñidas con hematoxilina-eosina para el análisis histológico y por inmunohistoquímica para la detección del CD44. Resultados: : Los cortes con hematoxilina-eosina mostraron tejido óseo con presencia de matriz osteoide y matriz ósea madura de aspecto usual. De las muestras CD44+, 80% fueron de aloinjerto y 20% de xenoinjerto. Las muestras con aloinjerto-xeoninjerto fueron negativas. No hubo diferencias en la intensidad de la expresión del CD44 entre las muestras positivas. El marcador se expresó en osteocitos, células estromales, infiltrado mononuclear y algunos histiocitos. El 80% de las muestras CD44+ y el 100% de las muestras con marcación de 60 o más células correspondían a aloinjertos (p=0,000). El 67% de las muestras del sector anterior y el 40% del sector posterior fueron CD44+ (p=0,689). Conclusión: Este estudio muestra por primera vez que la regeneración ósea guiada usando aloinjertos, es más eficiente para la generación de hueso maduro determinado por la expresión de CD44, comparado con el uso de xenoinjertos y mezcla de aloinjerto-xenoinjerto, independientemente del sector anatómico regenerado.


Subject(s)
Humans , Male , Female , Hyaluronan Receptors/metabolism , Alveolar Bone Grafting , Osteocytes , Bone Regeneration , Dental Implants , Hyaluronan Receptors/genetics , Allografts , Heterografts
4.
São Paulo; s.n; 2020. 59 p. ilust, tabelas.
Thesis in Portuguese | LILACS, Inca | ID: biblio-1178935

ABSTRACT

Introdução: O câncer de mama (CM) está associado ao aumento contínuo na incidência, com altas taxas de mortalidade em vários países. Existem quatro imunofenotipos de CM, luminal A, Luminal B, triplo negativo e HER2+. Algumas proteínas que estão presentes nas células tumorais e no microambiente tumoral fazem parte do processo da tumorigênese e metástases. A molécula de CD44 é considerada um importante marcador de resposta inflamatória e está presente em todos os leucócitos e na superfície de células tronco tumorais, estando envolvido com invasão tumoral e metástase. Alguns autores demonstraram que o signal transduce randactivator of transcription 3 (STAT3) e sex determining region Y related group box 2 (SOX2) estão relacionados com a regulação da divisão celular de células somáticas e com a tumorigênese e a metástase no CM. Objetivo: Avaliar a expressão de CD44+, SOX2 e STAT3 no sangue de pacientes com câncer de mama localmente avançado e metastático. Métodos: É um estudo de coorte transversal realizado no Hospital de Câncer de Pernambuco (HCP) e no Laboratório de Pesquisa Translacional do Instituto de Medicina Integral Prof. Fernando Figueira (IMIP) no período de março de 2017 a abril de 2018. Participaram deste estudo, 65 pacientes do sexo feminino com diagnóstico de câncer de mama e idade entre 28 a 64 anos. Destas, 51 mulheres tinham tumor de mama em estádio III (localmente avançado) e 14 com doença metastática (estágio IV). Como grupo controle, foram 24 mulheres clinicamente saudáveis com idade entre 18 anos a 65 anos. As análises da expressão de CD44+, SOX2 e STAT3 foi realizada por citometria de fluxo. Resultados: foram observados níveis elevados de células CD44+, CD24neg e CD44+/CD24neg CD44+/CD24negSOX2+ no sangue das pacientes com tumores de mama LB, HER2+ e Triplo negativos comparados aos controles (p<0,0001). Níveis baixos CD44+/CD24negSTAT3+ no sangue das pacientes comparado aos controles. Foram observados níveis elevados de células CD44+/CD24neg e CD44+/CD24negSTAT3+ no com CM localmente avançado comparado a CM metastático (p<0,05). As pacientes com CM triplo negativo apresentaram níveis elevados de linfócitos TCD3+SOX2+ comparado aos grupos com subtipos LB e HER2+ (p<0,05). O grupo de pacientes CM HER2+ apresentou baixos níveis de TCD3+SOX2+ comparado aos controles e LA (p<0,05). Níveis elevados de linfócitos TCD3 totais p=0,0007 e baixos de TCD3+SOX2+ p=0,02 no CM localmente avançado comparado a doença metastática. Níveis elevados de linfócitos TCD3+SOX2+ em pacientes linfonodos negativos quando comparado aos grupos N1 (p=0,004) e N2 (p<0,0001). Foram observados níveis elevados de células CD24neg e CD44+/CD24negSTAT3+ e de linfócitos TCD3+, e baixos níveis de TCD3+STAT3+ no CMTM localmente avançado comparado ao metastático. Conclusão: Os níveis das moléculas CD44, CD24, SOX2+ e STAT3+ no sangue mostraram ser alterados nas pacientes com CM em estágios mais avançados independente do imunofenótipo


Introduction: Breast cancer (BC) is associated with a continuous increase in incidence, with high mortality rates in several countries. There are four immunophenotypes of BC, luminal A, Luminal B, triple negative (TNBC), and HER2 +. Some proteins that are present in tumor cells, and the tumor microenvironment are part of the process of tumorigenesis and metastases. The CD44 molecule is considered an important marker of the inflammatory response. It is present in all leukocytes and on the surface of tumor stem cells, being involved with tumor invasion and metastasis. Some authors have demonstrated that the signal transducer and activator of transcription 3 (STAT3) and sex determining region Y related group box 2 (SOX2) are related to the regulation of somatic cell division and tumorigenesis and metastasis in BC. Objective: To evaluate the expression of CD44 +, SOX2, and STAT3 in the blood of patients with locally advanced and metastatic breast cancer. Methods: A cross-sectional study was carried out at the Pernambuco Cancer Hospital (HCP) and at the Translational Research Laboratory of the Instituto de Medicina Integral Prof. Fernando Figueira (IMIP) from March 2017 to April 2018. 65 female patients diagnosed with breast cancer participated in this study, aged between 28 and 64 years. Of these, 51 women had stage III (locally advanced) breast tumor, and 14 with metastatic disease (stage IV). As a control group, there were 24 clinically healthy women aged 18 to 65 years. The analysis of CD44 +, SOX2, and STAT3 expression was performed by flow cytometry. Results: elevated levels of CD44 +, CD24neg and CD44+/CD24neg, CD44+/CD24neg SOX2+ cells were observed in the blood of patients with LB, HER2+ and TNBC compared to controls (p<0.0001). Low CD44+/CD24negSTAT3+ levels in patients' blood compared to controls. Elevated levels of CD44+/CD24neg and CD44+/CD24negSTAT3 + cells were observed in locally advanced compared to metastatic BC (p <0.05). Patients with TNBC had elevated TCD3+SOX2+ lymphocytes compared to groups with subtypes LB and HER2+ (p <0.05). HER2+ patient had low levels of TCD3+ SOX2+ compared to controls and LA (p <0.05). High levels of total TCD3 (p= 0.0007) and low levels of TCD3 + SOX2 + (p = 0.02) in locally advanced BC compared to metastatic disease. Elevated levels of TCD3+SOX2+ in negative lymph node patients compared to groups N1 (p = 0.004) and N2 (p <0.0001). High levels of CD44+/CD24negSTAT3+ cells and TCD3+ were observed, and low levels of TCD3+ STAT3+ in locally advanced TNBC compared to metastatic. Conclusion: the levels of CD44, CD24, SOX2 +, and STAT3 + molecules in the blood were altered in breast cancer more advanced independent immunophenotype


Subject(s)
Humans , Female , Adult , Middle Aged , Aged , Transcription Factors , Breast Neoplasms , T-Lymphocytes , Hyaluronan Receptors , Flow Cytometry
5.
Journal of Experimental Hematology ; (6): 1794-1798, 2019.
Article in Chinese | WPRIM | ID: wpr-781395

ABSTRACT

OBJECTIVE@#To investigate the expression of CD44, CD87 and CD123 in acute leukemia and its correlation with cellular immune markers.@*METHODS@#A total of 166 patients with acute leukemia (AL) admitted from May 2014 to February 2017 were enrolled in AL groups. Among these patients, 100 patients suffered from acute myeloid leukemia, 50 patients suffered from acute lymphoid leukemia, and 16 patients showed B/medullary phenotype. At the same time 50 patients with non-acute leukemia were enrolled in the control group. 5 ml of fasting venous blood collected from the patients in each group, and the percentage of CD44, CD87 and CD123 cells was determined by three-color flow cytometry. Symptomatic chemotherapy was given to the patients with confirmed acute leukemia, and the remission was evaluated after 2 treatmen courses. The Complete remission (CR) was recorded and the percentage of CD44, CD87 and CD123 cells under different curative efficacy were recorded. The correlation of the prognosis patients with CD44, CD87 and CD123 was analyzed by SPSS Pearson correlation analysis software.@*RESULTS@#The positive rates of CD44, CD87 and CD123 in AL group were all higher than those in the control group (P<0. 05). The positive rates of CD44 and CD123 in acute myeloid leukemia group were higher than those in acute lymphoblastic leukemia group and B/myeloid phenotype group (P<0. 05). The positive rate of CD44 in acute lymphoid leukemia group was higher than that in B/medullary double phenotype group (P<0.05). The treatment in the patients of AL group was successfully completed. 132 patients reachel to CR and 34 patients to PR+NR after 2 courses. The positive rates of CD44, CD87 and CD123 in CR patients were lower than those in PR+NR patients (P<0.05). The results of SPSS Pearson correlation analysis showed that the prognosis of patients with acute leukemia negatively correlated with CD44 and CD87 (P<0.05).@*CONCLUSION@#The expression of CD44, CD87 and CD123 in different phenotype of acute leukemia are different, which correlateds with prognosis. The determination of CD44, CD87 and CD123 can be used to evaluate the prognosis of patients for the reference of clinical treatment.


Subject(s)
Humans , Hyaluronan Receptors , Allergy and Immunology , Immunity, Cellular , Interleukin-3 Receptor alpha Subunit , Allergy and Immunology , Leukemia, Myeloid, Acute , Prognosis , Receptors, Urokinase Plasminogen Activator , Allergy and Immunology
6.
Article in Chinese | WPRIM | ID: wpr-771928

ABSTRACT

OBJECTIVE@#To investigate the effects of low-dose decitabine on levels of soluble CD44 and GDF11, and hematopoietic function in elderly patients with myelodysplastic syndrome (MDS).@*METHODS@#Ninety-nine patients with senile myelodysplastic syndrome (MDS) admitted to our hospital from October 2015 to October 2017 were divided into group A, B and C according to their treatment, each with 33 cases.The patients in group A were treated with low-dose decitabine, the patients in group B were treated with usual dose of decitabine, and the patients in group C were treated with low-dose decitabine plus G-GSF, cytarabine, and aclarithromycin. The changes of soluble CD44, GDF11 levels and hematopoietic function (sTfR/E) were compared before and after treatment. The clinical remission rate and adverse reaction rate in 3 groups were analyzed.@*RESULTS@#Before treatment, the levels of CD44, GDF11 and sTfR/E were not significantly different between the 3 groups (P>0.05). After treatment, the levels of CD44 and GDF11 were significantly decreased in these groups, while the serum levels of sTfR/E were significantly increased, and there was no significant difference between the 3 groups (P>0.05). After treatment, the total effective rates of A, B, and C 3 group were 82.3%, 81.8%, and 78.8%, respectively, without statistically significant difference (P>0.05). During the treatment, the incidence of non-hemotoxic adverse reactions in group A was 8.8%, significantly lower than that in group B and C (30.3%, 27.3%) (P<0.05, P<0.05), the incidence of hemotoxic adverse reactions in group A was 39.4%, significantly lower than that 63.6% and 66.7% in group B and C (P<0.05, P<0.05).@*CONCLUSION@#Low-dose decitabine alone is effective in treating elderly patients with MDS as compared with conventional dose and combination therapy, moreover can significantly reduce the levels of CD44 and GDF11, improve hematopoietic function and low the adverse reactions. Thereby the low dose of decitabine may be a new choice for clinical treatment of MDS.


Subject(s)
Aged , Antineoplastic Combined Chemotherapy Protocols , Azacitidine , Bone Morphogenetic Proteins , Decitabine , Therapeutic Uses , Growth Differentiation Factors , Hematopoietic Stem Cell Transplantation , Humans , Hyaluronan Receptors , Myelodysplastic Syndromes , Drug Therapy , Treatment Outcome
7.
São Paulo; s.n; 2019. 99 p. ilust.
Thesis in Portuguese | LILACS, Inca | ID: biblio-1179955

ABSTRACT

Introdução: O câncer de pênis (CaPe) tem uma alta incidência em alguns países subdesenvolvidos. O tratamento padrão é a remoção do tumor primário e a linfadenectomia inguinal, em alguns pacientes. O fator prognóstico mais importante é a doença linfonodal, porém os métodos de estadiamento, disponíveis na atualidade são pouco precisos, e a alta taxa de morbidade da linfadenectomia tem estimulado o estudo de biomarcadores preditivos de metástases em linfonodos, selecionando os pacientes que necessitam da linfadenectomia. Os marcadores STAT3 e CD44, CD24, e SOX2+ são conhecidos por serem considerados marcadores de diagnóstico e prognóstico em outros cânceres, mas sem estudos no câncer de pênis. Objetivo: Avaliar os níveis das células CD44high/CD24low E TCD3+, e a expressão dos fatores de transcrição SOX2 e STAT3 no carcinoma escamoso de pênis. Métodos: Estudo transversal realizado no Hospital de Câncer de Pernambuco (HCP) e Laboratório de Pesquisa Translacional do Instituto de Medicina Integral Prof. Fernando Figueira (IMIP), no período de março de 2015 a dezembro de 2017 em uma população de 38 pacientes com CaPe e controles. Foram realizadas análises de SOX2, STAT3, CD24 e CD44 no sangue e tecido tumoral por citometria de fluxo. Resultados: A mediana da idade foi de 61 anos (37 - 80 anos). A maioria dos pacientes apresentou tumor localizado na glande (76,3%), tipo histológico usual (71,0%), de grau 1 (86,8%), sem invasão vascular e perineural (86,8% e 68,4%), sem presença de metástases (86,8%). Com relação ao estadiamento, 55,3% dos pacientes apresentavam estágio pT2 e 34,2% pT1. Foram encontrados níveis elevados de CD44highCD24low, CD44highCD24lowpSTAT3+ e CD44highCD24lowSOX2+nos pacientes, quando comparados aos controles (p<0,05). Foram observados níveis percentuais reduzidos de leucócitos SOX2+ e TCD3+/SOX2+, e elevados leucócitos pSTAT3 nos pacientes, quando comparados aos dos controles saudáveis. Foram constatados níveis elevados de CD44highCD24low e de leucócitos SOX2+ no grupo de pacientes com invasão perineural, tamanho tumoral > 3 cm e em estádio pT2 (p<0,05). Foram encontradas: associação dos níveis de célulasCD44high/CD24low e no sangue periférico com o tamanho do tumor (p=0,04) e status de sobrevida (p=0,01) e de CD44high/CD24lowSOX2+ no sangue e tumor com status de sobrevida nos pacientes;expressão reduzida de pSTAT3 no sangue de pacientes com invasão perineural e elevados no tecido tumoral no grupo em estádio pT1;alta expressão de TCD3+ no sangue e tecido tumoral de pacientes com tumor ≤3cm;aumento de TCD3+SOX2+ no sangue de pacientes sem invasão perineural e em estádio pT1.Observou-se correlação de CD44highCD24lowpSTAT3+ (r=0,669; p=0,024), de pSTAT3 (r=0,487; p=0,018) e de TCD3+SOX2+ (r=0,404; p=0,029) entre o sangue e o tecido tumoral. Conclusão: As moléculas CD44, CD24 e SOX2 foram marcadores de doença avançada, sendo associados ao pior prognóstico no câncer de pênis, porém, p-STAT3 e TCD3+ foram associados a um prognóstico mais favorável neste estudo


Introduction: Penile cancer (CaPe) has a high incidence in some underdeveloped countries. The standard treatment is removal of the primary tumor and inguinal lymphadenectomy in some patients. At the moment, the most important prognostic factor is lymph node disease, but the staging methods currently available are not very accurate and the high morbidity rate of lymphadenectomy has prompted the study of biomarkers that may predict lymph node metastasis, and aid in the selection of patients who need lymphadenectomy. The markers STAT3 and CD44, CD24, SOX2 + are known to be markers of diagnosis and prognosis in other cancers, but there are no such studies on penile cancer. Aim: To evaluate the levels of CD44high/ CD24low and TCD3+ cells, and the expression of SOX2 and STAT3 transcription factors in squamous cell carcinoma of the penis. Methods: A cross-sectional study was conducted at the Hospital de Cancer de Pernambuco (HCP) and Translational Research Laboratory at the Prof. Fernando Figueira Institute of Integral Medicine (IMIP), from March 2015 to December 2017 of a population of 38 patients with CaPe and controls. Analyses of SOX2, STAT3, CD24 and CD44 were performed on blood and tumor tissue by flow cytometry. Results: The median age was 61 years (37-80 years). Most of the patients presented tumor localized in the glans (76.3%), histological type (71.0%), grade 1 (86.8%), without vascular and perineural invasion (86.8% and 68.4% %), without metastases (86.8%). Regarding staging, 55.3% of the patients had pT2 and 34.2% pT1. High levels of CD44highCD24low, CD44highCD24lowpSTAT3+and CD44highCD24low,SOX2+ were found in the patients compared to the controls (p <0.05). Reduced percentage levels of SOX2+ and TCD3+/ SOX2+ leukocytes and high pSTAT3 leukocytes were observed in the patients when compared to healthy controls. There were elevated levels of CD44highCD24low and SOX2+ leukocytes in the group of patients with perineural invasion, tumor size> 3 cm and at pT2 stage (p <0.05). Peripheral blood CD44highCD24low levels were found to be associated with the tumor size(p = 0.04) and survival status (p = 0.01) and CD44highCD24lowSOX2+ levels in the blood and tumor with survival status in patients. Reduced levels of pSTAT3 in the blood of patients with perineural invasion and elevated tumor tissue in the pT1 stage. There were elevated levels of TCD3 + in the blood and tumor tissue of patients with tumors of ≤3cm. Increased TCD3 + SOX2 + was found in the blood of patients without perineural and pT1 stage invasion. CD44highCD24lowpSTAT3 + (r = 0.669; p = 0.024), pSTAT3 (r = 0.487, p = 0.018) and TCD3 + SOX2 + (r = 0.404, p = 0.029) correlations were observed between blood and tumor tissue. Conclusion: CD44, CD24 and SOX2 molecules were markers of advanced disease, and were associated with the worst prognosis in penile cancer. However, p-STAT3 and TCD3 + were associated with a more favorable prognosis in this study


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Penile Neoplasms , Transcription Factors , T-Lymphocytes , Hyaluronan Receptors , STAT3 Transcription Factor , Flow Cytometry
8.
Arq. bras. med. vet. zootec. (Online) ; 70(5): 1388-1396, set.-out. 2018. graf, ilus, tab
Article in English | ID: biblio-946856

ABSTRACT

This study aimed to evaluate the effect of the 0.15% sodium hyaluronate (SH) and of 0.5% carboxymethylcellulose (CMC) on tear film breakup time (TFBUT) in 10 healthy dogs and in 32 eyes of dogs with keratoconjunctivis sicca (KCS). In addition, the goblet cell density (GCD) of this population was quantified. TFBUT was assessed at baseline and at different time points following the instillation of SH and CMC. KCS was graded as mild, moderate, and severe. GCD were quantified from conjunctival biopsies. The number of GCD differed significantly between patients with mild and moderate KCS (P<0.01). TFBUT of healthy dogs increased only for 1 minute after treatment with SH (P<0.01). Regarding baseline and treatments, SH significantly increased TFBUT for up to 30 minutes on the ocular surface, in comparison to CMC, in all categories of KCS (P<0.01). TFBUT and GCD correlated positively when the healthy and diseased eyes were grouped (r=0.41, P=0.006). It can be concluded that in dogs with KCS, SH lasts longer periods on the ocular surface than CMC, but such agents does not increase TFBUT in healthy dogs. Additionally, tear film stability tends to reduce in a linear fashion from the mild to severe form of KCS.(AU)


Objetivou-se avaliar os efeitos do hialuronato de sódio a 0,15% (HS) e da carboximetilcelulose a 0,5% (CMC) no teste de ruptura do filme lacrimal (TRFL) em 10 cães saudáveis e em 32 olhos de cães com ceratoconjuntivite seca (CCS). Ademais, quantificou-se a densidade de células caliciformes (DCC) deles. Mensurou-se o TRFL em momentos distintos antes e após a instilação do HS e da CMC. Graduou-se a CCS em leve, moderada e severa. Quantificou-se a DCC a partir de biópsias conjuntivais. A DCC diferiu apenas entre pacientes com CCS leve e severa (P<0,01). Em cães saudáveis, o TRFL se elevou apenas após um minuto do tratamento com HS (P<0,01). Relativamente ao período basal e entre os tratamentos, o HS elevou o TRFL de forma mais eficaz e permaneceu por até 30 minutos na superfície ocular, comparativamente à CMC, em todas as categorias de CCS (P<0,01). Ao se agruparem os olhos saudáveis e os com CCS, o TRFL se correlacionou com a DCC (r=0.41, P=0.006). Conclui-se que o HS permanece por maior tempo na superfície ocular que a CMC em cães com CCS, mas que tais substâncias não elevam o TRFL em cães saudáveis. Ademais, a estabilidade do filme lacrimal tende a se reduzir de modo linear da forma leve até à severa da CCS.(AU)


Subject(s)
Animals , Dogs , Carboxymethylcellulose Sodium/adverse effects , Hyaluronan Receptors/analysis , Keratoconjunctivitis Sicca/veterinary
9.
Article in Chinese | WPRIM | ID: wpr-317604

ABSTRACT

<p><b>OBJECTIVE</b>To explore the predictive value of combination detection of Pgp1 expression in cancer tissue and serum CEA level for the prognosis of colorectal cancer (CRC) patients.</p><p><b>METHODS</b>Clinicopathological data, complete 5-year follow-up data and CRC tissue samples of 153 CRC patients with stage I( to II( tumor undergoing radical operation in our department from January 2004 to August 2006 were retrospectively collected. Immunohistochemical staining was used to detect the expression level of Pgp1. The combined evaluation of staining intensity and positive cell percentage was performed to determine the expression level of Pgp1. Pgp1 staining (-) and (+) was defined as low expression; and staining (++) and (+++) as high expression. Electrochemiluminescence immunoassay was used to detect the level of serum CEA. CEA > 5 μg/L was defined as positive. χand Fisher's exact test were performed to analyze the association of Pgp1 expression with CEA level and clinicopathological variables. Moreover, Kaplan-Meier method was used to analyze the survival. Univariate and multivariate Cox proportional hazard regression models were used to evaluate the roles of Pgp1 expression combined with serum CEA level in prognosis prediction.</p><p><b>RESULTS</b>Of 153 patients, 105 were males and 48 females with mean age of 59 (27 to 90) years; 41 cases were rectal cancer, and 112 cases colon cancer; 23 patients were TNM stage I( tumor, and 130 patients stage II( tumor; median follow-up time was 64 months; 30 cases were dead. Positive rate of Pgp1 expression in colorectal cancer tissues was 66.0%(101/153). The expression of Pgp1 was associated with gender, tumor location, and survival during the follow-up (all P<0.05). The preoperative positive rate of serum CEA was 28.1% (43/153). The preoperative serum CEA level was associated with tumor recurrence and survival (all P<0.05). Kaplan-Meier analysis showed the overall 5-year survival rate was 81.7%. The 5-year survival rate of patients with high expression of Pgp1 was 88.1%, which was significantly higher than 69.2% of those with low expression of Pgp1(P=0.003). The 5-year survival rate of patients with preoperative positive serum CEA was 72.1%, which was significantly lower than 86.1% of those with preoperative negative serum CEA(P=0.023). Furthermore, the 5-year survival rate of patients with negative Pgp1 plus positive CEA was 66.7%, which was significantly lower than 91.0% of those with positive Pgp1 plus negative CEA(P=0.002). Univariate analysis showed that gender, Pgp1 expression level, preoperative serum CEA level, and Pgp1 combined with CEA were significantly associated with the prognosis of patients(all P<0.05). Multivariate analysis showed that Pgp1 expression was an independent prognostic factor of CRC [HR(95%CI:1.261 to 64.224), P=0.028].</p><p><b>CONCLUSIONS</b>Low expression of Pgp1 in cancer tissue indicates poor prognosis in patients with stage I( and II( tumor. Combination detection of Pgp1 expression and serum CEA can be applied to predict the prognosis of patients with stage I( and II( colorectal cancer.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Biomarkers, Tumor , Blood , Physiology , Carcinoembryonic Antigen , Blood , Physiology , Colonic Neoplasms , Bodily Secretions , Colorectal Neoplasms , Bodily Secretions , Female , Fluorescent Antibody Technique , Humans , Hyaluronan Receptors , Metabolism , Kaplan-Meier Estimate , Male , Middle Aged , Multivariate Analysis , Neoplasm Proteins , Blood , Physiology , Neoplasm Recurrence, Local , Neoplasm Staging , Predictive Value of Tests , Prognosis , Proportional Hazards Models , Rectal Neoplasms , Bodily Secretions , Retrospective Studies , Sex Factors , Survival Rate
10.
Article in Chinese | WPRIM | ID: wpr-357516

ABSTRACT

<p><b>OBJECTIVE</b>This study aims to investigate the expression and relationship of CD44 and CD133 in normal oral mucosa, oral potentially malignant disorder (OPMD), and oral squamous cell carcinoma (OSCC). This work also analyzes the relationship between such expression and clinical factors. This study intends to evaluate the clinical value of using CD44 and CD133 as indices to evaluate the carcinogenic potential of OPMD.</p><p><b>METHODS</b>Clinical data from 60 patients with OPMD, 60 patients with OSCC, and 10 cases of normal oral mucosa were analyzed. Double immunohistochemical analysis was applied to investigate the expression of CD44 and CD133 in paraffin sections of normal oral mucosa, OPMD, and OSCC tissues. Subsequently, the relationships between such expression and clinical factors were analyzed.</p><p><b>RESULTS</b>The positive rates of CD44 expression in the normal oral mucosa, OPMD, and OSCC tissues were 100.00%, 96.67%, and 71.67% (P<0.05), respectively. Meanwhile, the positive rates of CD133 expression in the normal oral mucosa, OPMD, and OSCC tissues were 0.00%, 35.00%, and 63.33% (P<0.05), respectively. The expression of CD44 and CD133 was found to be correlated (P<0.05). Such expression was related to the clinical stages and lymphatic metastasis of OSCC (P<0.05).</p><p><b>CONCLUSIONS</b>CD44 and CD133 can be used individually as clinical indices to evaluate the carcinogenic potential of OPMD.
.</p>


Subject(s)
AC133 Antigen , Carcinogenesis , Carcinoma, Squamous Cell , Humans , Hyaluronan Receptors , Lymphatic Metastasis , Mouth Mucosa , Mouth Neoplasms
11.
Neuroendocrinology ; 104(2): 183-193, 2017.
Article in English | LILACS, SES-SP, SESSP-IALPROD, SES-SP, SESSP-IALACERVO | ID: biblio-1024761

ABSTRACT

Background/Aims: Although craniopharyngioma (CP) is histologically benign, it is a pituitary tumour that grows rapidly and often recurs. Adamantinomatous CP (ACP) was associated with an activating mutation in ß-catenin, and it has been postulated that pituitary stem cells might play a role in oncogenesis in human ACP. Stem cells have also been identified in pituitary adenoma. Our aim was to characterize the expression pattern of ABCG2, CD44, DLL4, NANOG, NOTCH2, POU5F1/OCT4, SOX2, and SOX9 stem cell markers in human ACP and pituitary adenoma. Methods and Results: We studied 33 patients (9 ACP and 24 adenoma) using real-time quantitative PCR (RT-qPCR) and immunohistochemistry. SOX9 was up-regulated in ACP, exhibiting positive immunostaining in the epithelium and stroma, with the highest expression in patients with recurrence. CD44 was overexpressed in ACP as confirmed by immunohistochemistry. SOX2 did not significantly differ among the tumour types. The RT-qPCR array showed an increased expression of MKI67,OCT4/POU5F1, and DLL4 in all tumours. NANOG was decreased in ACP. ABCG2 was down-regulated in most of the tumours. NOTCH2 was significantly decreased in the adenomas. Conclusion: Our results confirm the presence of stem cell markers in human pituitary tumours as well as the different expression patterns of ACP and adenoma. These findings suggest that ACP may originate from a more undifferentiated cell cluster. Additionally, SOX9 immunodetection in the stroma and the highest expression levels related to the relapse of patients suggest a contribution to the aggressive behaviour and high recurrence of this tumour type.


Subject(s)
Pituitary Neoplasms/metabolism , Aged , Humans , Biomarkers, Tumor/metabolism , Adenoma/metabolism , Adenoma/pathology , Gene Expression , Child , Child, Preschool , Adolescent , Hyaluronan Receptors/metabolism , Craniopharyngioma/metabolism , Craniopharyngioma/pathology , Neural Stem Cells/metabolism
12.
Bauru; s.n; 2017. 112 p. tab, ilus, graf.
Thesis in Portuguese | LILACS, BBO | ID: biblio-880412

ABSTRACT

O objetivo deste estudo consistiu em avaliar a expressão imuno-histoquímica da podoplanina e do CD44v6 pelas células malignas, verificando a associação destas proteínas com as variáveis clínicas, microscópicas, com o índice histopatológico de malignidade e com a sobrevivência livre de doença de 91 pacientes portadores de carcinomas espinocelulares (CEC) de lábio inferior, tratados no Centro de Tratamento e Pesquisa do Hospital do Câncer A.C.Camargo, São Paulo. Os tumores foram corados, separadamente, com os anticorpos anti-podoplanina e anti-CD44v6, sendo avaliada a imunoexpressão destas proteínas pelas células neoplásicas, no front de invasão tumoral, por meio de um método semi-quantitativo de escores. A associação da expressão da podoplanina e do CD44v6 com as variáveis demográficas, clínicas e microscópicas foi feita pelo teste do qui-quadrado ou exato de Fisher. As taxas de sobrevivência livre de doença, acumuladas em cinco e dez anos, foram calculadas pelo teste de Kaplan-Meier e a influência das variáveis clínicas e microscópicas no prognóstico avaliadas pelo modelo de regressão de Cox. A correlação entre a podoplanina e o CD44v6 foi analisada pelo teste de Spearman. Em todos os testes estatísticos utilizou-se um nível de significância de 5%. Os resultados mostraram uma predominância da forte expressão membranosa e citoplasmática da podoplanina pelas células malignas. Verificou-se uma associação significativa da podoplanina citoplasmática com a recidiva locorregional (p=0,028) e da podoplanina membranosa com o índice histopatológico de malignidade tumoral (p=0,026). O CD44v6 foi fortemente expresso pelas células neoplásicas de 95,4% dos CECs e significativamente, associado com o estadiamento clínico T (p=0,034). Não houve correlação entre a podoplanina e o CD44v6 nos CECs de lábio inferior. A forte expressão de podoplanina membranosa (p=0,016) e citoplasmática (p=0,030) pelas células malignas foi fator de prognóstico favorável independente na sobrevivência livre de doença. Concluímos que a podoplanina e o CD44v6 são fortemente expressos pelas células neoplásicas e que a forte imunoexpressão membranosa e citoplasmática da podoplanina pode auxiliar na identificação do risco de recidiva locorregional nos pacientes portadores de carcinoma espinocelular de lábio inferior.(AU)


The aim of this study was evalute the podoplanin and CD44v6 immunohistochemical expression by malignant cells and its association with the clinical and microscopic variables, tumor histopathological grading and disease-free survival of 91 patients with lip squamous cell carcinomas (SCC), submitted to surgical treatment at Research and Treatment Center of the Cancer Hospital A.C. Camargo, São Paulo. The tumors were stained separately, with the antibodies anti-podoplanin and anti-CD44v6, and the immunoexpression of these proteins, by the neoplastic cells in the invasion front, was evaluated by a semi-quantitative scores method. Chi-square test or Fishers exact test was used to analyze the association of podoplanin and CD44v6 expression with demographic, clinical, and microscopic variables. Disease-free survival in five and ten years, were calculated by the Kaplan-Meier method and the influence of clinical and microscopic variables on prognosis were evaluated by the Cox regression model. The correlation between podoplanin and CD44v6 expression was analyzed by Spearman's test and a significance level of 5% was used in all statistical tests. The results showed a predominance of strong membranous and cytoplasmic podoplanin expression by malignant cells. An association between cytoplasmic podoplanin and locorregional recurrence (p=0,028) and membranous podoplanin with tumor histopathological grading (p=0,026). CD44v6 was strongly expressed in 95.4% of the SCCs neoplastic cells and significantly associated with the clinical staging T (p=0,034). There was no correlation between podoplanin and CD44v6 expression in the lower lip SCC. The strong expression of membranous (p=0.016) and cytoplasmic (p=0.030) podoplanin by malignant cells was a favorable independent prognostic factor in disease-free survival. Concluding, the podoplanin and CD44v6 are strongly expressed by neoplastic cells and the strong membranous and cytoplasmic immunoexpression of podoplanin can help the identification of locoregional recurrence risk in patients with squamous cell carcinoma of the lower lip.(AU)


Subject(s)
Humans , Male , Female , Middle Aged , Aged , Carcinoma, Squamous Cell/pathology , Hyaluronan Receptors/analysis , Lip Neoplasms/pathology , Membrane Glycoproteins/analysis , Neoplasm Recurrence, Local/pathology , Age Factors , Biomarkers, Tumor/analysis , Immunohistochemistry , Kaplan-Meier Estimate , Prognosis , Sex Factors , Statistics, Nonparametric
13.
Braz. j. med. biol. res ; 50(8): e6538, 2017. tab, graf
Article in English | LILACS | ID: biblio-888984

ABSTRACT

Cancer stem cells reside in a distinct region within the tumor microenvironment that it is believed to play a fundamental role in regulating stemness, proliferation, survival, and metabolism of cancer cells. This study aimed to analyze the effect of extracellular alkalinization on metabolism and survival of human CD24-/CD44+ breast cancer stem cells (BCSCs). BCSCs were cultured in alkalinized DMEM-F12 and incubated at 37°C, 5% CO2, and 20% O2 for 30 min, 6, 24, and 48 h. After each incubation period, we analyzed the modulation of various mRNA expressions related to pH and cellular metabolic regulation using the qRT-PCR. Metabolic state was measured using colorimetric and fluorometric assays. To examine cell proliferation and apoptosis, we used trypan blue and annexin V/propidium iodide assay, respectively. This study demonstrated that alkalinization could stimulate extracellular carbonic anhydrase (CAe) activity, as well as CA9 and HIF1α expression. Under alkaline pH and HIF1α regulation, glucose consumption, extracellular lactate production, and LDH activity of BCSCs were upregulated while O2 consumption was downregulated. These metabolic shifts seemed to promote apoptosis and suppress the proliferation of BCSCs. To conclude, modulation of the extracellular environment through alkalinization could change the metabolic states of BCSCs, which in turn affect the cell survival.


Subject(s)
Humans , Female , Breast Neoplasms/metabolism , CD24 Antigen/metabolism , Hyaluronan Receptors/metabolism , Neoplastic Stem Cells/metabolism , Apoptosis , Cell Proliferation , Cell Survival , Extracellular Space , Gene Expression Regulation, Neoplastic , Reverse Transcriptase Polymerase Chain Reaction
14.
Int. j. morphol ; 34(4): 1197-1202, Dec. 2016. ilus
Article in English | LILACS | ID: biblio-840866

ABSTRACT

Cancer has been considered as a stem cell disease. Suspension culture combined with anti-cancer drugs has recently been proposed for isolation of cancer stem cells (CSCs). In the current study, Vincristine as an anti-cancer drug combined with suspension culture was used for isolation and purification of CSCs from human breast cancer cell line (MDA-MB231). The cells were treated with different concentrations of vincristine (0, 2, 4, 6 and 8 ng/ml). Stem cells were identified with the expression of OCT4, nanog, SOX2 and nucleostemin genes by RT-PCR. Mammosphere forming unit was measured upon suspension culture containing EGF, bFGF, LIF, B27, insulin and BSA. The isolated mammospheres were investigated for CD44 expression. Results showed that 4 ng/ml of vincristine for 72 hours could be utilized as the best and most reliable dose which eliminates around 80 % of non-cancer stem cells with no destructive effect on CSCs' viability (P> 0.05). RT-PCR demonstrated that drug treated cells expressed OCT4, nanog, SOX2 and nucleostemin. Mammosphere formation unit of cells pretreated with vincristine was significantly higher than unpretreated ones (P>0.05). Immunofluorescence staining for CD44 depicted high expression of CSC marker among the isolated mammospheres. Vincristine combined with suspension culture can be considered as an appropriate method to isolate CSC.


El cáncer ha sido considerado como una enfermedad de células madre. Recientemente se ha propuesto cultivo en suspensión en combinación con medicamentos contra el cáncer para aislamiento de las células madre del cáncer (CMC). En este estudio se utilizó la vincristina como fármaco anticanceroso combinado con cultivo en suspensión para el aislamiento y purificación de las células madre cancerosas, de la línea celular de cáncer de mama humano (MDA-MB231). Las células se trataron con diferentes concentraciones de vincristina (0, 2, 4, 6 y 8 ng/ml). Las células madre se identificaron mediante la expresión de los genes OCT4, Nanog, SOX2 y nucleostemin por RT-PCR. La unidad de formación mammosphere se midió a través de cultivo en suspensión que contenía EGF, bFGF, LIF, B27, insulina y BSA. Los mammospheres aislados fueron estudiados para la expresión de CD44. Los resultados mostraron que 4 ng/ml de vincristina durante 72 horas podrían ser utilizados como la mejor y más fiable dosis que permite eliminar alrededor del 80 % de las células madre no cancerosas, sin causar un efecto destructivo sobre la viabilidad de las CMC (P> 0,05). La RT-PCR mostró que en las células tratadas con él fármaco hubo expresión de los genes OCT4, Nanog, SOX2 y nucleostemin. La unidad de formación de las células pretratadas con vincristina fue significativamente más alta que las unidades sin tratamiento previo (P>0,05). La inmunofluorescencia para CD44 muestró una alta expresión del marcador de CMC entre mammospheres aisladas. La vincristina en combinación con el cultivo en suspensión puede ser considerado como un método apropiado para aislar CMC.


Subject(s)
Humans , Female , Antineoplastic Agents, Phytogenic/pharmacology , Breast Neoplasms/pathology , Neoplastic Stem Cells/drug effects , Neoplastic Stem Cells/pathology , Vincristine/pharmacology , Hyaluronan Receptors/metabolism , Cell Line, Tumor , Fluorescent Antibody Technique, Indirect , Nanog Homeobox Protein , Octamer Transcription Factor-3/genetics , Octamer Transcription Factor-3/metabolism , Reverse Transcriptase Polymerase Chain Reaction , SOXB1 Transcription Factors/genetics , SOXB1 Transcription Factors/metabolism , Staining and Labeling , Tumor Stem Cell Assay
15.
Article in English | WPRIM | ID: wpr-277914

ABSTRACT

Objective To investigate the expressions of CD44,CD47,and c-met in ovarian clear cell carcinoma (OCCC) tissue and their correlations with clinical variables and prognosis. Methods Immunohistochemical method was used to investigate the expressions of CD44,CD47,and c-met in tissues from 86 OCCC patients and the relationships of their expressions with the clinicopathological factors of OCCC were analyzed. Results The expressions of CD44,CD47,and c-met were significantly high in OCCC tissues (90.7%,91.9%,and 94.2%,respectively). The strong positive expressions of CD44 and CD47 were significantly correlated with advanced International Federation of Gynecology and Obstetrics stages,chemotherapeutic resistance,and poor prognosis (all P<0.05),the strong positive expression of c-met was significantly correlated with chemotherapeutic resistance and poor prognosis (all P<0.05),whereas there was no correlation between the strong positive expressions of CD44,CD47,and c-met and the lymphatic node metastasis. COX survival analysis revealed that advanced International Federation of Gynecology and Obstetrics stages and high expressions of CD44,CD47 and c-met were independent risk factors for poor prognosis (P<0.05). There was a positive correlation between CD44 (or CD47) and c-met and between CD44 and CD47 (the Spearman correlation coefficient rwas 0.783,0.776,and 0.835,respectively,all P<0.01). Conclusions The expressions of CD44,CD47,and c-met increase in OCCC tissues and are correlated with each other. High expressions of CD44,CD47,and c-met are independent factors for poor prognosis.


Subject(s)
Adenocarcinoma, Clear Cell , Metabolism , CD47 Antigen , Metabolism , Female , Humans , Hyaluronan Receptors , Metabolism , Lymphatic Metastasis , Ovarian Neoplasms , Metabolism , Prognosis , Proto-Oncogene Proteins c-met , Metabolism , Survival Analysis
16.
Fortaleza; s.n; 2016. 68 p. ilus, tab.
Thesis in Portuguese | LILACS | ID: biblio-971952

ABSTRACT

O câncer gástrico é a quarta neoplasia mais comum em todo o mundo, representando a segunda causa de mortalidade por câncer. Apesar do tratamento com cirurgia e quimioterapia, a sobrevida global em cinco anos de pacientes com câncer gástrico permanece baixa. Uma possível explicação para ineficácia da terapia é a presença de células-tronco cancerosas, uma subpopulação de células tumorais que apresentam características de células-tronco. Estas células, assim como as células tronco embrionárias, são consideradas imortais, podem se auto-renovar e se transformar em qualquer célula do corpo. Vários marcadores, incluindo CD44 e CD133, têm sido relatados como marcadores de células-tronco, normais e cancerosas, e utilizados para isolar células cancerosas de tumores sólidos. O objetivo desse trabalho foi avaliar a expressão de CD44 e de CD133, no câncer gástrico primário e metástases linfonodais, através de imunohistoquímica, e relacioná-la com as variáveis clínico-patológicas de tipo histológico, sexo, idade, localização anatômica, dimensão do tumor, invasão angiolinfática, infiltração perineural, classificação TNM (TN) e acometimento linfonodal. Este estudo foi desenvolvido a partir de um conjunto de 72 casos de adenocarcinoma gástrico, dos Arquivos do Serviço de Patologia e Medicina Legal da Universidade Federal do Ceará (DPML-UFC). Utilizou-se a técnica de tissue microarray asociada à imunohistoquímica comanticorpo monoclonal anti-CD44 e policlonal anti-CD133...


Gastric cancer is the fourth most common cancer worldwide, accounting for the second leading cause of cancer mortality. Despite treatment with surgery and chemotherapy, the overall five-year survival of patients with gastric cancer remains low. One possible explanation for the ineffectiveness of therapy is the presence of cancer stem cells, a subpopulation of tumor cells that have stem cell characteristics. It has been reported that these, as well as embryonic stem cells, are immortal, can self-renew and to differentiate to be transformed in any cell type in the body. Several markers, including CD44 and CD133, have been reported as stem cell markers in both normal and cancerous cells and have been used to isolate cancer cells from solid tumors. The aim of this study was to evaluate the expression of CD44 and CD133 in primary gastric cancer and lymph node metastases by immunohistochemical and to relate it to clinicopathologic variables as histological type, gender, age, anatomical site, tumor size, angiolymphatic invasion, infiltration perineural, TNM classification (TN) and lymph node involvement. This study was developed from a set of 72 cases of gastric adenocarcinoma, from the Archives of Pathology and Forensic Medicine of the Federal University of Ceará Service (DPML-UFC). Tissue microarray andimmunohistochemistry were utilized, with anti-CD44 monoclonal antibody and polyclonal anti-CD133...


Subject(s)
Humans , Stomach Neoplasms , Stem Cells , Hyaluronan Receptors
17.
Iranian Journal of Cancer Prevention. 2015; 8 (5): 27-34
in English | IMEMR | ID: emr-175777

ABSTRACT

Background: Recent evidence has suggested that epithelial cancers including colorectal cancer [CRC] have driven by a small population of self-renewing, multi-potent cells termed cancer stem cells [CSCs] which could be responsible for recurrence of cancer. Aldehyde dehydrogenase 1 [ALDH1] activity has used as a functional stem cell biomarker to isolate CSCs in different cancers such as colorectal cancer


Objectives: The main aim of this research was to determine the utility of ALDH1 activity along with CD44 and EPCAM in identifying stem cell-like cells in human HT-29 colonic adenocarcinoma cell line


Materials and Methods: In this experimental study, colon CSCs biomarkers including CD44, EPCAM and ALDH1 in colonospheres and parent cells have analyzed by flow cytometry. The expression levels of stemness genes in spheroid and parental cells have investigated using SYBR Green real-time PCR. In addition, in vivo xenografts assay has performed to determine tumorigenic potential of tumor spheroid cells in nude mice


Results: According to results, over 92% of spheroids were CD44+/EpCAM+, while parent cells only have expressed 38% of CD44/EpCAM biomarkers [P < 0.001]. Controversially, ALDH activity was about 2-fold higher in the parent cells than spheroid cells [P < 0.05]. In comparison with the parental cells, expression levels of ''stemness'' genes, like Sox2, Oct4, Nanog, C-myc, and Klf4 have significantly increased in colonosphere cells [P < 0.05]. Further, administration of 2500 spheroids could be sufficient to initiate tumor growth in nude mice, while 1x106 of parental cells has needed to form tumor


Conclusions: For the first time, we have shown that colonospheres with low ALDH1 activity has indicated increased tumorigenic potential and stemness properties. So, it hasn't seemed that ALDH1 could become a useful biomarker to identify CSCs population in HT-29 cell line


Subject(s)
Humans , Aldehyde Dehydrogenase , Isoenzymes , Retinal Dehydrogenase , Biomarkers, Tumor , Neoplastic Stem Cells , HT29 Cells , Hyaluronan Receptors , Antigens, Neoplasm , Cell Adhesion Molecules
18.
Article in Chinese | WPRIM | ID: wpr-239154

ABSTRACT

<p><b>OBJECTIVE</b>To explore the role of CD44 in monocyte adhesion to human brain microvascular endothelial cells (HBMECs) and monocyte migration across an in vitro model of blood-brain barrier (BBB) infected by Cryptococcus neoformans (Cn).</p><p><b>METHODS</b>An in vitro blood-brain barrier model was constructed using a transwell chamber covered with a HBMEC monolayer. The wild-type strain of Cn B4500FO2, TYCC645#32 strain with CPS1 gene deletion and PCIP strain with CPS1 complementation were chosen to infect the monolayer HBMECs. THP-1 cells were added to the upper chamber of transwell, and the relative migration rate was determined by counting the number of the cells entering the lower chambers. The inhibitory effects of anti-CD44 monoclonal antibody and the CD44 inhibitor bikunin were examined on THP-1 binding to and migration across HBMECs.</p><p><b>RESULTS</b>Cn infection of the HBMECs caused markedly enhanced THP-1 cell adhesion and migration across the monolyers (P<0.01) dependent on Cn concentration and exposure time. Addition of anti-CD44 monoclonal antibody and bikunin significantly lowered THP-1 adhesion and migration rates in the BBB model with Cn-infected HBMECs (P<0.01) with a dose dependence of the antibody (within 0-1 µg) and inhibitor (within 0-20 nmol/L). Both THP-1 adhesion rate and migration rate were lowered in the BBB model infected with CPS1 gene-deleted Cn but increased in the model infected with the complemented strain compared with those in the wild-type strain-infected model.</p><p><b>CONCLUSION</b>In the in vitro BBB model, CD44 expressed on HBMECs may play an essential role in monocyte adhesion to and migration across the BBB. The capsular hyaluronic acid may mediate Cn-induced monocyte adhesion and migration.</p>


Subject(s)
Blood-Brain Barrier , Allergy and Immunology , Microbiology , Brain , Cell Biology , Microbiology , Cell Line , Cryptococcosis , Allergy and Immunology , Cryptococcus neoformans , Endothelial Cells , Microbiology , Humans , Hyaluronan Receptors , Metabolism , Monocytes , Cell Biology
19.
Article in Chinese | WPRIM | ID: wpr-288022

ABSTRACT

OBJECTIVE To explore downstream regulatory pathway of microRNA-21 (miR-21) in colon cancer cells (RKO) through detecting miR-21 and its target PDCD4, and the influence of miR-21 regulation on the sensitivity of RKO cells to 5-fluorouracil (5-FU). METHODS 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the effect of 5-FU on the viability of RKO cells with knockout of miR-21 or high expression of PDCD4. Real-time was used to determine the expression of PDCD4, ABCC5 and CD44 in RKO cell after knockout of miR-21. RESULTS MTT assay reveals that the IC50 of 5-FU in RKO-WT cells (52.82 ± 0.06 umol/L) was about 67% higher than in miR-21 knockout cells (32.23 ± 0.05 umol/L) (P < 0.05), and the apoptosis ratio elevated after knockout of miR-21. High expression of PDCD4, a target gene of miR-21, can negatively regulate the expression of ABC transporter ABCC5 and the stem cell marker CD44. CONCLUSION MiR-21 can mediate the drug resistance to 5-FU by inhibiting its target PDCD4, which can regulate the expression of ABCC5 and CD44 genes.


Subject(s)
ATP Binding Cassette Transporter, Subfamily G, Member 5 , ATP-Binding Cassette Transporters , Genetics , Antimetabolites, Antineoplastic , Pharmacology , Apoptosis Regulatory Proteins , Physiology , Cell Line, Tumor , Colonic Neoplasms , Drug Therapy , Pathology , Drug Resistance, Neoplasm , Fluorouracil , Pharmacology , Humans , Hyaluronan Receptors , Genetics , Lipoproteins , Genetics , MicroRNAs , Physiology , RNA-Binding Proteins , Physiology
20.
Chinese Journal of Stomatology ; (12): 675-680, 2015.
Article in Chinese | WPRIM | ID: wpr-294648

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effect of co-culture system of bone marrow mesenchymal stem cells (BMSC) and vascular endothelial cells (EC) on osteogenesis.</p><p><b>METHODS</b>BMSC were isolated by whole bone marrow centrifugal adherent method. Then BMSC were induced into EC with induced medium. Co-culture system in different proportions of BMSC and EC (10:0, 10:1, 8:2, 7:3, 5:5, 3:7, 2:8, 1:10, 0:10) were further evaluated. The cell growth level of BMSC was examined. The CD44 expression of BMSC and von willebrand factor (vWF) expression of vascular EC were examined by immunofluorescence. Furthermore, calcium nodules exhibited by alizarin red staining, alkaline phosphatase activity, and the expression of osteogenic genes by reverse transcription-quantitative PCR (RT-qPCR) were observed to validate the osteogenesis of co-culture system.</p><p><b>RESULTS</b>The growth curve of P3 passage of BMSC demonstrated the doubling time of BMSC was 39.9 h. The positive specific markers of BMSC and EC showed efficient induction. Although the calcium nodules ratio of the co-culture [group 7:3 (19.0 ± 3.0) and group 5:5 (20.8 ± 2.9)] was not significantly different (P > 0.05), but higher than that of other co-culture groups with a significant difference (P < 0.01). Alkaline phosphatase activity was increased with prolonged induction of osteogenic medium. While alkaline phosphatase activity of group 10:0 (16.84 ± 0.82), group 10:1 (15.86 ± 3.10), group 8:2 (16.37 ± 1.33), group 7:3 (17.99 ± 1.98), and group 5:5 (17.49 ± 0.87) did not show significant difference after osteogenic induction for 7 days (P > 0.05), but significantly higher than that of other co-culture groups (P < 0.05). The co-culture ratio of 7:3 (33.74 ± 0.99) was slightly higher than that of 5:5 (31.09 ± 0.87), but significantly higher than that of other groups (P < 0.01). Moreover, the osteocalcin (OCN) and runt-related transcription factor 2 (RUNX2) expression of group 7:3 was significantly higher than that of other groups.</p><p><b>CONCLUSIONS</b>The EC that derived from BMSC can promote the BMSC differentiate into osteoblasts. The co-culture system of BMSC and EC with the ratio of 7:3 increases the alkaline phosphatase activity and facilitates the expression of osteogenic genes.</p>


Subject(s)
Alkaline Phosphatase , Metabolism , Bone Marrow Cells , Cell Biology , Physiology , Cell Culture Techniques , Cell Differentiation , Cell Proliferation , Coculture Techniques , Core Binding Factor Alpha 1 Subunit , Metabolism , Endothelial Cells , Cell Biology , Physiology , Humans , Hyaluronan Receptors , Metabolism , Mesenchymal Stem Cells , Cell Biology , Physiology , Osteoblasts , Cell Biology , Osteocalcin , Metabolism , Osteogenesis , Physiology , Time Factors , von Willebrand Factor , Metabolism
SELECTION OF CITATIONS
SEARCH DETAIL