Association of FAT atypical cadherin 1 with clinicopathological parameters and prognosis in esophageal squamous cell carcinoma / 中华消化外科杂志

Objective:To investigate the association of FAT atypical cadherin 1 (FAT1) with clinicopathological parameters and prognosis in esophageal squamous cell carcinoma (ESCC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 124 patients with ESCC who were admitted to Shanxi Cancer Hospital from January 2011 to December 2015 were collected. There were 85 males and 39 females, aged from 40 to 72 years, with a median age of 60 years. The ESCC tissues surgically removed and adjacent tissues specimens were collected to prepare tissue microarray for immunohistochemical staining. The 5 cases of ESCC tissues and adjacent tissues were analyzed by real-time quantitative polymerase chain reaction (qRT-PCR). Observation indicators: (1) the expression of FAT1 protein in ESCC and adjacent tissues; (2) the expression of FAT1 RNA in ESCC and adjacent tissues; (3) the expression of FAT1 protein in ESCC tissues and its association with clinicopathological parameters; (4) follow-up and survival. Follow-up using outpatient examination and telephone interview was conducted to detect survival of patients up to February 13, 2019. The survival time was from surgical date to tumor-related death or endpoint of follow-up. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was analyzed using the non parameter rank sum test. The Kaplan-Meier method was used to calculate survival time, and Log-rank test was used for survival analysis. Results:(1) The expression of FAT1 protein in ESCC and adjacent tissues: of 124 specimens, the 107 cases of ESCC tissues and 93 cases of adjacent tissues were finally obtained because of exfoliative tissues. There were 76 cases of ESCC tissues and corresponding adjacent tissues matched. Results of immuno-histochemical staining showed that FAT1 protein was expressed in both ESCC and adjacent tissues and was brown after staining. FAT1 was located in cytomembrane, with high expression of FAT1 as ≥75 and low expression as <75. The relative expression levels of FAT1 protein in ESCC and adjacent tissues were 68±42 and 77±37, showing a significant difference between ESCC and adjacent tissues ( t=2.380, P<0.05). (2) The expression of FAT1 RNA in ESCC and adjacent tissues: results of qRT-PCR showed that the relative expression levels of FAT1 RNA in 5 cases of ESCC and adjacent tissues were 1.6±0.4 and 2.5±0.3, with a significant difference between them ( t=3.560, P<0.05). (3) The expression of FAT1 protein in ESCC tissues and its association with clinicopathological parameters: of the 107 ESCC patients, 58 cases had high expression of FAT1. There were 42 and 16 cases with high expression of FAT1 in 65 non-drinking patients and 42 drinking patients, respectively, showing a significant difference between them ( χ2=7.229, P<0.05). (4) Follow-up and survival: 96 of 107 ESCC patients were followed up for 38.0?94.9 months, with a median follow-up time of 45.9 months. Survival analysis showed that the survival time of patients with high FAT1 expression was 24 months, versus 22 months of patients with low FAT1 expression, indicating no significant difference between them ( χ2=1.773, P>0.05). Results of subgroup analysis showed that the survival time was 24 months and 21 months of female patients with high and low FAT1 expression, 23 months and 22 months of non-smoking patients with high FAT1 expression and low FAT1 expression, 23 months and 21 months of non-drinking patients with high FAT1 expression and low FAT1 expression, respectively, showing significant differences between them ( χ2=8.769, 12.827, 10.724, P<0.05). Conclusions:The expression of FAT1 in ESCC tissues is low. Female, non-smoking and non-drinking ESCC patients with high FAT1 expression have good survival.

Avaliação de marcadores de prognóstico no câncer de mama e análise funcional de CIP4 / Evaluation of prognostic markers in breast cancer and functional analysis of CIP4

O câncer de mama é uma doença extremamente heterogênea compreendendo diferentes subtipos moleculares que resultam em evoluções clínicas e condutas terapêuticas distintas. A maior gravidade desta patologia está associada a sua capacidade de formação de metástases Mudanças no padrão de expressão gênica têm sido associadas à manifestação do fenótipo metastático. Neste trabalho, utilizamos microarranjos de tecido (TMAs) para investigar a expressão de 8 biomarcadores candidatos (CIP4, PPIL1, ITGAV, AKAP14, MICA, FXYD1, ARPC3, ABG1) e avaliar seu potencial prognóstico em pacientes com carcinoma ductal invasivo da mama. Destes, ARPC3 PPIL1 e CIP4 mostraram associações estatisticamente significativas com a sobrevida câncer específica e/ou a probabilidade de desenvolvimento de metástases. Determinamos que a expressão aumentada de CIP4 nos tumores está associada a maior probabilidade de desenvolvimento de metástases. CIP4 é uma proteína adaptadora descrita na literatura como moduladora de migração e invasão celular e portanto selecionamos este candidato para caracterização funcional detalhada. Observamos que a expressão de CIP4 encontra-se aumentada em linhagens tumorais com características invasivas. A partir do silenciamento estável e regulado de CIP4 na linhagem metastática MDA-MB-231, determinamos que CIP4 modula positivamente a ativação de MAPK-p38 e a expressão de MMP2 , sugerindo que CIP4 participe em vias de sinalização importantes para a transição epitélio-mesenquima (EMT). O silenciamento de CIP4 resultou em uma redução de aproximadamente 50% da capacidade migratória e invasiva das células tumorais in vitro , e na diminuição da formação de metástases pulmonares in vivo. Coletivamente, nossos resultados indicam que CIP4 tem potencial como marcador de prognóstico assim como um possível alvo terapêutico no controle da disseminação de metástases nos tumores da mama

Breast cancer is an extremely heterogeneous disease comprising different molecular subtypes that result in different clinical outcomes and therapeutic procedures. The severity of this disease is mainly associated with its ability to produce metastasis. Changes in gene expression profile have been associated with the manifestation of the metastatic phenotype. In this study, we used tissue microarrays (TMAs) to investigate the expression of 8 candidate biomarkers (CIP4, PPIL1, ITGAV, AKAP14, MICA, FXYD1, ARPC3 e ABG1) and to evaluate their prognostic potential in patients with invasive ductal breast carcinoma. Among these, ARPC3, PPIL1 and CIP4 showed statistically significant associations with cancer specific survival and/or the patient's probability to develop metastasis. We found that increased expression of CIP4 in tumors is associated with a higher probability of developing metastasis. CIP4 is an adaptor protein described in the literature as a modulator of cell migration and invasion and therefore we selected this candidate for detailed functional characterization. We observed that CIP4 expression is increased in tumor cell lines with invasive characteristics. Following the stable and regulated knockdown of CIP4 in the metastatic line MDA-MB-231, we determined that it modulates positively the activation of MAPK-p38 and the expression of MMP2, suggesting that CIP4 participates in important signaling pathways required for the epithelial mesenchymal transition (EMT). CIP4 silencing resulted in an approximate 50% reduction of the migratory and invasive capacity of tumor cells in vitro and decreased the generation of lung metastases in vivo. Collectively, our results indicate that CIP4 has potential as a prognostic marker as well as a potential therapeutic target to control the metastatic dissemination of breast tumors

Quick and inexpensive method to elaborate tissue punches useful in paraffin tissue microarrays / Método rápido y económico para la elaboración de dispositivos de punch útiles en microarreglos de tejidos en parafina

The tissue microarrays (TMAs) were first called multitumor block. In 1998 was described the current technique, that uses an innovated sampling method for more than 1,000 cylindrical paraffin tissue core biopsies in a single paraffin block. TMAs are now considered as a useful powerful research tool in Histology and Pathology laboratories, for the standardization of immunohistochemical techniques along with in situ hybridization. However, one disadvantage to its widespread use is the high cost of professional paraffin tissue punches, and the complexity in the development of homemade devices previously described in other studies. This study describes a step by step process to develop four different home-made devices made with materials that are common in hospitals and offices. These devices are useful in Histopathology laboratories to obtain paraffin blocks with until 360 samples of tissue, investing from two to fifteen dollars in the development of each device described.

Los microarreglos de tejido (TMAs) fueron llamados por primera vez como bloque multitumor. En 1998 se describió la técnica actual, que utiliza un novedoso método de muestreo para obtener más de 1,000 cilindros de biopsias de tejidos incluidos en un solo bloque de parafina. Actualmente, los TMAs se consideran una poderosa herramienta de investigación en laboratorios de Histología y Patología, para la estandarización de técnicas inmunohistoquímica e hibridación in situ entre otras. Sin embargo, uno de los inconvenientes para su uso generalizado es el alto costo de los dispositivos profesionales para tejidos en parafina, y la complejidad en la elaboración de los dispositivos caseros descritos previamente en otros estudios. Este estudio describe paso a paso el proceso de elaboración de cuatro dispositivos caseros útiles para la obtención de matrices de tejido elaborados con materiales que son comunes en hospitales y oficinas. Estos dispositivos son útiles en laboratorios de Histopatología con el fin de obtener bloques de parafina de hasta 360 muestras de tejido, con una inversión de 2 a 15 dólares en la elaboración de cada uno de los dispositivos descritos.

Estudo de candidatos a biomarcadores moleculares de prognóstico em carcinoma renal de células claras / Study of molecular biomarker candidates for prognosis in clear cell renal carcinoma

O carcinoma de células renais (CCR) é o tumor mais agressivo que afeta o rim de pessoas adultas. O CCR é uma doença heterogênea, com diferentes alterações moleculares e variados patrões histológicos e clínicos que apresentam evolução diferente. Atualmente apenas variáveis anatomopatológicas clássicas são utilizadas para determinar o prognóstico dos pacientes. Utilizando uma plataforma de microarranjos de DNA, nosso grupo identificou em um trabalho anterior um conjunto de genes que se encontram diferencialmente expressos em tumores de rim. Neste estudo, nove candidatos foram selecionados para avaliação como marcadores de prognóstico no CCR. Foi confirmada a alteração na expressão dos genes ARNTL, ACTN4 e EPAS1 (p < 0,05) em amostras tumorais de CCR através de PCR em tempo real. Adicionalmente, foi observada a alteração da expressão dos genes ARNTL, EPAS1 e CASP7 em linhagens celulares imortalizadas derivadas de tumores renais, recapitulando por tanto, as alterações observadas nos tumores obtidos de pacientes. Posteriormente investigamos o padrão de expressão proteica destes candidatos por imunohistoquímica utilizando microarranjos de tecidos. Foi detectada a diminuição significativa (p < 0,05) da expressão das proteínas ACTN4, ARNTL, CASP7 e EPAS1 em tumores de pacientes com CCR relativamente ao tecido renal não tumoral. Além disso, foi possível determinar valores de imunomarcação capazes de estratificar pacientes com CCR em diferentes grupos de risco quanto à sobrevida câncer-específica, que adicionalmente apresentaram associação significativa com parâmetros anatomopatológicos utilizados na clínica. As imunomarcações de ACTN4, ARNTL, e EPAS1 se mostraram parâmetros independentes de prognóstico de sobrevida dos pacientes. A imunomarcação de CASP7 foi capaz de identificar subgrupos de pacientes com pior prognóstico dentro de um conjunto de pacientes de baixo risco em função do estadio clinico, além de identificar pacientes com menor risco de morte pelo câncer entre aqueles apresentaram recorrência em até 5 após a cirurgia. O conjunto de resultados obtidos aponta para um novo conjunto de biomarcadores moleculares com potencial relevância para auxiliar no prognóstico de pacientes com carcinoma de células renais

The renal cell carcinoma (RCC) is the most aggressive tumor that affects the kidney in adult people. The RCC is a heterogeneous disease, with many different molecular alterations and varied histological and clinical patterns with different outcome. Currently, only classic anatomopathological variables are used to determine patients' prognosis. Using a DNA microarray platform, our group identified in a previous work a set of genes differentially expressed in renal tumors. In this study, nine candidates were selected for evaluation as prognostic biomarkers in RCC. Alteration of the gene expression in RCC tumor samples was confirmed for ARNTL, ACTN4 and EPAS1 (p < 0.05) by real time PCR. Additionally, gene expression changes of ARNTL, EPAS1 and CASP7 were also observed in immortalized cell lines derived from renal tumors, recapitulating the expression changes detected in the patients' tumors. Next, we used tissue microarrays to investigate the protein expression of the selected candidates by immunohistochemistry. Expression of the proteins ACTN4, ARNTL, CASP7 and EPAS1 was detected as significantly downregulated (p < 0.05) in patients´ tumors relative to non-tumor renal tissue. Furthermore, immunostaining patterns of the selected candidates were able to stratify patients with RCC in different risk groups according to cancer-specific survival, which also showed significant associations with anatomopathological parameters used in the clinics. ACTN4, ARNTL and EPAS1 immunostaining resulted as independent prognostic parameters of patient survival. CASP7 immunostaining was able to identify subgroups of patients with worse prognosis in a set of low risk patients as determined by their clinical stage, and also identified patients with lower risk of death from cancer amongst patients that relapsed within 5 years after surgery. Overall, these results point to a new set of molecular biomarkers with potential relevance to help in the prognosis of patients with renal cell carcinoma

The Expression of PIK3CA in Gastric Adenocarcinoma / 天津医药

Objective To investigate the relationship between phosphatidylinositol 3-kinase catalytic subunit α(PIK3CA) expression and the incidence and different pathological grade of gastric cancer, and the mechanism thereof. Meth-ods The expressions of PIK3CA, serine/threonine protein kinase (pAkt) and cell proliferation associated nuclear antigen (ki-67) in gastric carcinoma and adjacent tissues and normal gastric mucosa were detected by immunohistochemical method. The relationship between expressions of PIK3CA, pAkt and ki-67 and tumorigenesis was analyzed. The expressions of PIK3CA, pAkt and ki-67 in different pathological conditions of gastric tissues were analyzed. The relationship between tu-mor pathologic classification and differentiation were analyzed too. Results There were significantly higher expressions of PIK3CA, pAkt and ki-67 in gastric cancer (P＜0.05), which were the highest in the poorly differentiated gastric adenocarci-noma (P＜0.05). There were a positive correlation between expressions of PIK3CA, pAkt and ki-67 and different pathologi-cal levels of gastric carcinoma and adjacent tissues and normal gastric tissues (P＜0.05). Conclusion PIK3CA may be the initiating factor of PI3K/Akt signaling pathway, which induced phosphorylation of Akt and activation PI3K/Akt signaling pathway, promoting the proliferation, invasion and metastasis of gastric adenocarcinoma.

A novel tissue microarray instrumentation:The HT-1 tissue microarrayer.

Background: Tissue microarray (TMA) is a novel and useful tool to efficiently analyze gene expression in histological tissues. Aim: Cost-efficient and easy to use automated tissue arrayers will provide a better instrumentation to generate TMAs. Thus, we designed and produced our tissue microarrayer to meet these needs. Materials and Methods: The HT-1 tissue microarrayer we designed and manufactured consists primarily of four parts, including an instrument to make array pores for the recipient paraffin blocks, a punch needle, an instrument for negative-pressure embedding, and a special manipulator. By using the HT-1, 14 different TMAs were made to accommodate 312 cases of tissues and TMA sections were tested by hematoxylin-eosin (H&E) staining, in situ hybridization, and immunohistochemistry. Results: Expand: Hematoxylin and eosin staining showed that the tissue cylinders were similar, even, and in order on the slides. Most importantly, the HT-1 microarrayer can make array pores in the recipient paraffin block with a single application in seconds. The HT-1 also contains a unique negative pressure system for embedding TMA blocks. In addition, HT-1 can make tissue cylinders with the same levels and depth for equally embedded and sectioning. Conclusions: The HT-1 tissue microarrayer is a device that is simple, economical and easy to use.

Androgen receptor levels during progression of prostate cancer in the transgenic adenocarcinoma of mouse prostate model.

Aim To construct tissue microarrays (TMAs) that consisted of prostate tumours from the transgenic adenocarcinoma of mouse prostate (TRAMP) mice and non-transgenic murine prostates and to assess androgen receptor (AR) levels during progression of prostate cancer in TRAMP mice by immunohistochemistry. Methods Haematoxylin and eosin (H&E) sections from the ventral and dorso-lateral prostate lobes of non-transgenic, intact TRAMP and castrated TRAMP were used to demarcate regions of interest for TMAs construction. The samples on TMAs were used to evaluate AR expression using video image analysis (VIA). Results AR was expressed during cancer progression, but AR levels were reduced or absent in late stage disease. Furthermore, when AR levels were compared in tumours from intact and castrate animals, a significant increase in AR levels was observed following androgen ablation. Conclusion Similar to clinical prostate cancer, in the TRAMP model, prostate tumours evolve mechanisms to maintain AR expression and AR responsive gene pathways following castration to facilitate continued tumour growth.

Application of Tissue Microarrays to Study Protecting Effects of Baicalin and Octreotide on Lung Injury in Severe Acute Pancreatitis / 医学研究杂志

Objective To observe the influence of Baicalin and Octreotide on lung injury of rat with severe acute pancreatitis(SAP) and discuss the therapeutic effect and mechanism of the two medicines on SAP.Methods The improved Aho(?)method was adopted to pre- pare SAP rat models via retrograde injection of 3.5% sodium taurocholate to the pancreatic duct.The 135 SAP rat models after being pre- pared were randomly divided into the model group,Baicalin treatment group and Octreotide treatment group with 45 rats in each group;an- other 45 were selected to be the sham operation group which only received abdomen opening surgery.The above - mentioned groups were then randomly divided into 3h,6h and 12h groups with 15 rats in each group.Observed respectively at 3h,6h and 12h after operation,the mortalities of all rat groups followed by batch execution of rats,and then observed the gross pathological and pathological changes of lung. The tissue microarrays technology was applied to prepare the lung tissue microarrays sections.The changes in Bax and bcl-2 protein ex- pression levels of lung tissue of each group were observed via immunohistochemical staining and meanwhile the TUNEL method was applied to observe the lung cell category and apoptotic index changes of lung tissue in each group.Results The 12h survival of model group was 66,67% while those of Baicalin treatment group and Octreotide treatment groups were both 100%,indicating a marked difference(P＜0.05).Comparison of lung pathological score disclosed:The model group and Baicalin treatment group obviously exceeded the sham oper- ation group at various time points(P＜0.01);at 6h and 12h the Baicalin treatment group was obviously less than the model group(P＜0.05)and the Oetreotide treatment group less than the model group(P＜0.01).Comparison of Bax protein of lung in each group dis- closed:The Baicalin treatment group and Octreotide treatment group obviously exceeded the sham operation group and model group at 3h (P＜0.05),meanwhile the Baicalin treatment group obviously exceeded the Octreotide treatment group(P＜0.01).Changes of lung bcl -2 protein expression level of all groups disclosed:The model group and Baicalin treatment group obviously exceeding the sham operation group(P＜0.01)as well as the Octreotide treatment group(P＜0.05)at different time points,the Baicalin treatment group was obviously less than the model group at 6h(P＜0.05).The peak of apoptosis occurred to the Baicalin treatment group at 6h,but statistics showed no obvious difference among all groups at different time points(P＞0.05).Conclusions(1)Both Baicalin and Octreotide have protecting effect on SAP lung injury and their therapeutic mechanism is possibly related to apoptosis.The application of tissue microarrays in SAP pathological examination can save manpower and material resources,cut down the experimental cost and improve the experimental efficien- cy,thus worth popularizing.

Significance of the Expression of p27(Kip1) Protein in Human Breast Cancer / 한국유방암학회지

PURPOSE: p27(Kip1) is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. It binds to a variety of cyclin/CDK complexes, inhibits kinase activity and blocks the cell cycle as a negative regulator. Reduced p27(Kip1) expression has been reported to be a significant predictor of poor survival in numerous human breast cancers. We executed p27(Kip1) protein assay in primary breast cancer and compared these result with known prognostic parameters. METHODS: Immunohistochemical assay was performed on tissue microarrays from 183 patients with breast cancer to evaluate the biologic and clinical significance of p27(Kip1) expression. RESULTS: Decreased p27(Kip1) expression was significantly associated with clinical stage (P=0.027), low nuclear grade (P<0.001), high histologic grade (P<0.001), high score mitotic index (P<0.001), increased Ki67 labeled index (P=0.006) and negative estrogen receptor status (P=0.0024). In survival analysis, p27(Kip1) was useful to predict disease- free survival (P=0.0106) and overall survival (P=0.0154) of the patient after surgery followed by chemotherapy or radiation therapy. CONCLUSION: Reduced expression of p27(Kip1) protein was as sociated with biologically aggressive phenotype of breast cancer and was an useful to predict patient outcome.

Apoptosis Related Protein Expressions in Immunohistochemical Staining Using Tissue Mi croarrays of Breast Cancer

PURPOSE: In order to confirm the clinical application of a tissue microarrays method, the expression rate and relationship between factors related apoptosis, hormonal receptors and the clinical factors were investigated. METHODS: A tissue microarrays of 59 breast cancer tissues, and apoptosis related factors were examined by immunohistochemical staining using monoclonal antibodies. RESULTS: The median age of the patients was 49.9 years and 86.4% had a pathological stage of over stage II. The average number of metastatic lymph nodes was 3.8. p53 expression was noted in 21 cases (35.6%) and was related to Bcl-2, ER and PR expression. PTEN was expressed in 39 cases (66.1%) and related to FAS, Bcl-2, ER and PR expression. Fas was expressed in 34 cases (57.6%) and related with PR and BAX expression. BAX expression was observed in 42 cases (71.2%) and was related to the metastatic axillary lymph nodes, and both Bcl-2 and PR expression. Bcl-2 expression was noted in 33 cases (55.9%) and related to ER and PR expression. ER was expressed in 34 cases (57.6%) and was related positively with PR expression. CONCLUSION: The tissue microarrays method can be used for both screening and analyzing many factors or different tumor types. This new technique may be very powerful for the rapid identification of the tumor characteristics.

KAI1/CD82 protein expression in human hepatocellular carcinoma and its relationship with invasion and metastasis / 中国普通外科杂志

ObjectiveTo investigate the relationship between KAI1/CD82 protein expression in human hepatocellular carcinoma (HCC) and invasion and metastasis.MethodsSpecimens from primary hepatocellular carcinoma (155 cases), tumorous emboli (22 cases), intrahepatic satellite metastases (4 cases), extrahepatic metastases (16 cases), and normal livers (5 cases) for which a tissue microarray was constructed and used for immunohistochemical detection of KAI1/CD82 protein expression. ResultsImmunohistochemical analysis of tissue microarrays demonstrated KAI1/CD82 protein expression in 61%(95/155)of the primary HCCs and only in 32%(7/22) of the tumorous emboli ( P

Apoptosis Related Protein Expressions in Immunohistochemical Staining Using Tissue Microarrays of Breast Cancer / 한국유방암학회지

PURPOSE: In order to confirm the clinical application of a tissue microarrays method, the expression rate and relationship between factors related apoptosis, hormonal receptors and the clinical factors were investigated. METHODS: A tissue microarrays of 59 breast cancer tissues, and apoptosis related factors were examined by immunohistochemical staining using monoclonal antibodies. RESULTS: The median age of the patients was 49.9 years and 86.4% had a pathological stage of over stage II. The average number of metastatic lymph nodes was 3.8. p53 expression was noted in 21 cases (35.6%) and was related to Bcl-2, ER and PR expression. PTEN was expressed in 39 cases (66.1%) and related to FAS, Bcl-2, ER and PR expression. Fas was expressed in 34 cases (57.6%) and related with PR and BAX expression. BAX expression was observed in 42 cases (71.2%) and was related to the metastatic axillary lymph nodes, and both Bcl-2 and PR expression. Bcl-2 expression was noted in 33 cases (55.9%) and related to ER and PR expression. ER was expressed in 34 cases (57.6%) and was related positively with PR expression. CONCLUSION: The tissue microarrays method can be used for both screening and analyzing many factors or different tumor types. This new technique may be very powerful for the rapid identification of the tumor characteristics.