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Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.
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Objective To study the relationship of microRNA-145(miR-145) and CD44 in the hepatocellular carcinoma(HCC).Methods 13 clinical samples of HCC tissues and corresponding normal liver tissues were collected at department of General Surgery,Second Affiliated Hospital of Fujian Medical University from October 2015 to June 2016.The patients with positive expression of CD44 were studied.The expression levels of miR-145 and CD44 in HCC tissues and corresponding normal tissues were detected by real-time quantitative PCR.Western blot was performed to detect the expression of CD44 in HCC cells Hep G2 and SNU423.Biological information methods predicted whether miR-145 and CD44 have a targeted relationship.CD44 expression levels were detected after high expression of miR-145 in HCC cell line with positive expression of CD44.The vector and luciferase reporter genes were constructed to verify the interaction between miR-145 and CD44.The effects of miR-145 on proliferation in HCC cell lines with positive and negative CD44 expression were examined by tetramethylazoazole salt (MTT) assay.Results 8 of the 13 patients showed positive CD44 expression in HCC tissues.Compared with normal liver tissues,the relative expression of miR-145 in HCC tissues was significantly lower (0.998±0.010 vs.0.503±0.046,P＜0.05),and the relative expression of CD44 was higher (0.996±0.005 vs.1.878±0.108,P＜0.05).Bioinformatics suggested that miR-145 had a targeted relationship with CD44.High expression of miR-145 can significantly reduce the expression level of CD44 mRNA in HCC cell SNU423 (0.941±0.010 vs.0.515±0.021,P＜0.05) and down-regulate the expression of CD44 protein.Confirmed by luciferase reporter assay,CD44 is the target gene of miR-145.After transfection with miR-145 mimics,the proliferation of CD44 positive cell SNU423 was significantly inhibited (P＜0.05).Conclusions miR-145 can affect the proliferation of CD44 positive HCC cells by regulating the expression of CD44,which may be one of the pathogenesis of HCC.
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Objective To investigate the expression of CD133 and CD44 proteins in gastric stromal tumors (GST) and their clinical significances. Methods The expression of CD44 and CD133 proteins in the GST tissues of 112 patients was detected by immunohistochemical staining. The relation between the expression of CD44 and CD133 proteins and the clinicopathological characters was analyzed. The survival and prognosis of GST were also analyzed. Results Both CD44 and CD133 were expressed on the cell membranes. The expression rates of CD44 and CD133 were 58.04 % (65/112) and 42.86 % (48/112) separately; the co-expression rate of CD44 and CD133 was 27.68 % (31/112). CD44 and CD133 were negative in normal peritumoral tissues. No correlation was found between CD44 and CD133 and the clinicopathological parameters including gender, age and lymphatic vessel invasion (all P>0.05), but the expression levels of CD44 and CD133 in patients with the mitotic count ≥ 5/50 high-power field, large diameter and vascular invasion were significantly higher (all P0.05), but the co-expression level of CD44 and CD133 in patients with tumor diameter ≥5 cm was significantly higher than that in patients with tumor diameter < 5 cm (χ2=5.040, P=0.025). The overall survival rate of the patients with co-expression of CD44 and CD133 was shorter than that in other groups (χ2 = 8.758, P= 0.001). No correlation was found between CD44 and CD133 expression (r=0.126, P=0.210). Multivariate analysis with the Cox regression models showed that the tumor diameter ≥5 cm (P=0.042) and co-expression of CD44 and CD133 (P=0.003) were significantly associated with poor prognosis. Conclusion CD44 and CD133 as robust cancer stem cell markers in GST might be the prognostic factors.
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Objective To construct an overexpression lentiviral vector of CD44 and obtain an androgen-dependent prostate cancer LNCaP cell line with CD44 overexpression.Methods CD44 gene sequence was obtained by PCR amplification.The synthesized oligonucleotides were cloned in the lentiviral vector LV5 after digested with NotI and NsiI enzyme.Recombinant plasmids were verified by enzyme digestion analysis and sequencing,and were transfected into 293T package cells.Supernatant was collected and concentrated to obtain lentivirus solution at a high titer.The titer of virus was identified by multiple proportion dilution methods.Then the recombinant viruses were transfected into LNCaP cells.Results Fluorescence intensity in infected LNCaP cells was increased,detected by a fluorescence microscope.And QRT PCR and Western blot results showed that the expression of CD44 was significantly increased(P＜0.05).Conclusions The overexpression lentiviral vector of CD44 is successfully constructed,and the LNCaP cell line with CD44 overexpression is obtained.
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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)
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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, NonparametricABSTRACT
BACKGROUND:Effective sorting of prostate cancer stem cels is the basis of experimental studies in prostate cancer developing. The most common sorting method is magnetic-activated cel sorting. OBJECTIVE:To separate CD133+/CD44+ cels in prostate cancer tissues using self-designed magnetic beads folowed by culture, passage and immunological identification. METHODS:Self-designed magnetic microspheres were applied to establish immunomagnetic beads to sort CD133+/CD44+ cels in prostate cancer tissues. The sorted cels were cultured in serum-free medium. The sphere formation, cel morphology, and proliferation ability after cel passage were statisticaly compared between the sorted cels and the normal tumor cel lines. Immunofluorescence detection was performed to detect the expression of specific antibodies. RESULTS AND CONCLUSION:Self-designed immunomagnetic beads had smal diameter and a high-sorting effect. The sorted cels possessed a high capacity of microsphere formation. After cel culture and passage, the cels highly expressed CD133 and CD44 antigens. The sorted cels with no induction had varying shapes and grew vigorously. After induction with transforming growth factor-β, the cultured cels were noted to have a single shape and grow slowly. The cel proliferation ability of sorted cels in these two groups differed significantly from that of the normal cancer cel lines (bothP < 0.05). In conclusion, the CD133+/CD44+ cels sorted from prostate tumor cels possessed cel morphology and function characteristics of stem cels which can provide a basis for extraction and culture of prostate cancer stem cels. Cite this article:Gong R, Li SY, Huo ZX, Ding H, Sun EL. Extraction of prostate cancer stem cels using self-designed magnetic beads. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):36-41.
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Objective To investigate the expressions and clinical significance of survivin, and lymphatic vessel endothelial hyaluronan receptor 1 ( LYVE-1) in gastric cancers.Methods The expres-sions of survivin and LYVE-1 were detected with streptavidin avidin-peroxidase ( SP) immunohistochemical method in 70 cases of gastric cancers and 50 cases of normal gastric tissues.The relationship between the expressions of survivin and LYVE-1 with the clinicopathological parameters was analyzed.Results The ex-pression rate of survivin was 65.71%(46/70) in gastric cancers, and 6.00% (3/50) in normal gastric tissues, with statistically significant difference between two groups (χ2 =43.048, P 0.05).The expression rate of LYVE-1 was 32.86%(23/70) in gastric cancers, and 4.00%(2/50) in gastric ulcers, with significant difference between two groups (χ2 =14.726, P 0.05 ) .There was a positive correlation between expressions of survivin and LYVE-1 in gastric cancers ( r =0.271, P <0.05).Conclusions Survivin and LYVE-1 were abnormally expressed in gastric cancers with a colose relationships between two parameters.They might play important roles in tumorigenesis and progression of a gastric cancer.
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Objective To detect the expression of stem-cell related factors Nanog and CD44 in spheroid body-form?ing cells of gastric cancer cell line MKN-45. Methods The gastric cancer cell line MKN-45 was used to culture spheroid bodies in non-adherent condition in a serum-free medium supplemented with epidermal growth factor (EGF) and basic fibro?blast growth factor (bFGF). Using Western blot analysis, immunofluorescence staining and quantitative real time polymerase chain reaction(qRT-PCR), the expression levels of stem cell-related genes Nanog and CD44 were studied. Results In this study, we observed that MKN45 cells formed spheroid bodies in non-adherent condition in a serum-free medium, and the levels of Nanog and CD44 mRNA expression in spheroid body-forming cells were 2.34 ± 0.22 and 1.18 ± 0.04,respectively, which were higher than those in parental cells (1.00±0.00 and 1.00±0.05). The levels of Nanog and CD44 protein expression in spheroid body-forming cells were 0.18±0.02 and 0.24±0.04, respectively, which were significantly higher than those in pa?rental cells (0.07±0.02 and 0.18±0.01, P<0.05). Nanog protein was positively stained within the perinuclear and cytoplasm of the spheroid body-forming cells, and CD44 was positively stained mainly in the membrane. Dual staining of Nanog/CD44 indicated that the embryonal protein Nanog was co-localized with CD44 in the spheroid body-forming cells. Conclusion Spheroid body-forming cells developed from human gastric cancer cell line MKN-45 in serum-free medium supplemented with EGF and bFGF show characteristics of cancer stem cell (CSC). The cells co-expressed of CD44 and Nanog maybe a phe?notype of gastric CSCs.
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BACKGROUND:Tumor stem cels not only initiate tumorigenesis, but also are involved in the invasion and metastasis of tumor cels. For tumor stem cels, to identify the specific cel surface marker has become a research hotspot. OBJECTIVE:To explore the clinical significance of cancer stem cel surface marker CD44 in gastric cancer invasion and lymph node metastasis. METHODS: CD44 protein expression in specimens of gastric cancer tissue was detected by the immunohistochemical SABC method. The relationship between CD44 protein expression and biological characteristics and prognosis of gastric cancer was detected using Pearsonχ2 test and Cox regression analysis. RESULTS AND CONCLUSION:Among 100 cases of gastric carcinoma, 59 cases (59%) were positive for CD44 protein expression. CD44 protein expression in normal gastric mucosa at above 5 cm from the edge of primary gastric cancer was negative. CD44 protein was widely expressed in tissues of gastric cancer, mainly expressed in the cel membrane, and a smal amount of expression in the cytoplasm. CD44 protein expression in gastric cancer tissue was not correlated with sex of the patients or age (P > 0.05), but was associated with tumor staging and lymph duct tissue infiltration, histological grade, and tumor size (P < 0.05). Deep tumor invasion, high histological grade, big diameter of tumor, and lymph node metastasis could lead to high positive CD44 protein expression. Positive expression of CD44 is an independent prognostic factor affecting postoperative survival (P < 0.05). The results show that the cancer stem cel surface markers CD44 in gastric carcinoma tissues is strongly associated with invasion of gastric carcinoma and lymph node metastasis. High expression of CD44 presents poor prognosis.
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Objective To investigate the expressions of CD133 and CD44 and their prognostic significance in gastrointestinal stromal tumors (GIST).Methods Streptavidin perosidase (SP) method of immunohistochemistry was used to detect expressions of CD133 and CD44 proteins in 42 cases of GIST, and the relationship between their expressions and tumor size, mitotic count were analyzed by univariate and multivariate factor analyses.Results The expressions of CD133 and CD44 proteins in GIST were 21.4％ (9/42) and 78.6％ (33/42), respectively.The expressions of CD133 and CD44 proteins were significantly correlated with tumor size and mitotic count (P ＜ 0.05).Univariate factor analysis showed that the overall survival of GIST patients with positive CD133 protein (23.2 months) was shorter than that of patients with negative CD133 protein(63.1 months) (P ＜ 0.05).The overall survival of GIST patients with negative CD44 protein (23.2 months) was shorter than that of patients with positive CD44 protein (63.3 months) (P ＜ 0.05).Multivariate factor analysis showed that tumor size, mitotic count and CD44 protein were independent prognostic indicators for survival time after operation.Conclusions The positive expressions of CD133 and CD44 proteins might be the prognostic factors of GIST patients.
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Objectives To observe the pre and post-operational changes of the expressions of survivin, caspase-3 and CD44V6 in patients with colorectal cancer after intra-abdominal implantation of sustained releasing fluorouracil. Meth-ods Sixty-four patients with colorectal cancer (Dukes’stage of B and C) were divided into treatment group and control group, 32 patients in each group. The standard radical surgery was performed in two groups of patients. The fluorouracil im-plants were implanted intra-abdominally in treatment group. The peripheral blood levels of surviving and caspase-3 were de-tected by RT-PCR. The level of CD44V6 was detected by flow cytometry in two groups of patients. Results There were no significant differences in levels of survivin, caspase-3 and CD44V6 before surgery between two groups (P>0.05). The level of survivin (0.362 ± 0.183) was significantly lower at 14 days after operation in treatment group than that of control group (0.585±0.207), but the level of caspase-3 (2.001±0.146) was significantly higher than that of control group (1.654±0.111). The levels of CD44V6 were significantly lower in treatment group (1.857±0.535) and control group (3.471±0.496) after opera-tion than those before operation (9.557±1.170 and 9.729±0.943, P＜0.05), and the level of CD44V6 was significantly lower in treatment group than that of control group (P＜0.05). Conclusion The implant for the sustained release of fluorouracil showed a positive impact on micrometastases and prognosis of colorectal cancer, while improved the long-term efficacy of postoperative colorectal cancer.
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Objective To investigate the expression of CXCR7 in gastric carcinoma tissues and the role of CXCR7 in tumor growth, adhesion and invasion of gastric cancer. Methods The expression levels of CXCR7, Survivin, CD44v6 and matrix metalloproteinase (MMP)-3 were detected by immunnohistochemistry method in gastric cancer specimens came from 160 patients (gastric cancer group) and 30 specimens of normal gastric tissues (control group). Results The expression of CXCR7 protein was significantly higher in gastric cancer group than that of control group (P<0.05). The positive CXCR7 staining was significantly different with tumor size, depth of invasion, lymph node metastasis and clinical staging ( P<0.05). CXCR7 expression was positively correlated with Survivin, CD44v6 and MMP-3. Conclusion CXCR7 might take part in progression of gastric cancer by anti-apoptosis (Survivin). CXCR7 might take part in the lymph node metastasis of gastric cancer and mediate the adhesion and invasion by CD44v6 and MMP-3.
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Objective To explore the expressions and significances of sICAM-1 and sCD44v6 in peritoneal lavage fluid after laparoscopically assisted radical gastrectomy,in order to evaluate the risk of laparoscopically assisted radical gastrectomy for the peritoneal plant transfer.Methods Ninety-two patients with gastric cancer were randomly divided into the control group and the experimental group.The control group was treated with open laparotomy and the experimental group was treated with laparoscopic surgery.The concentrations of both sICAM-1 and sCD44v6 were detected by enzyme linked immunosorbent assay.The positive rates of tumor cells in the two groups were compared.Results The expressions of sICAM-1,sCD44v6 in the two groups before operation had no significant difference (t =0.198,P =0.543 ; t =0.897,P =0.429).After operation,sICAM-1 and sCD44v6 levels increased significantly,but there was no significant difference between the two groups (t =0.766,P =0.312 ; t =1.092,P =0.129).The positive rates of tumor cells in the two groups before and after operation have no significant differences (x2 =0.789,P =0.287 ; x2 =0.912,P =0.198).Conclusion Laparoscopically assisted radical gastrectomy does not increase the expression of sICAM-1 and sCD44v6 in peritoneal lavage fluid and the risk of peritoneal metastasis.It is worthy of clinical poptdarization and application.
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Objective To investigate the role ot adhesion molecule CD44 in respiratory syncytial virus (RSV) infection induced bronchiolitis.Methods Thirty-six children with RSV infection induced bronchiolitis (infection A group),34 children with only RSV infection (infection B group) and 40 healthy children (control group) were selected.The proportion of peripheral blood CD44+ cell was determined by flow cytometry.The levels of peripheral blood interferon (IFN)-γ,interleukin (IL)-4,IL-10 and IL-12 were determined by enzyme-linked immunosorbent assay.Results The expression of CD44 and the level of IL-4 in peripheral blood in infection A group were significantly higher than those in infection B group and control group,there were statistical differences (P ＜ 0.05).The level of IFN-γ in peripheral blood in infection A group was significantly lower than that in infection B group and control group,there were statistical differences (P ＜0.05).The levels of IL-12 in peripheral blood in infection A group and infection B group were significantly lower than that in control group,there were statistical differences (P ＜ 0.05).There were no statistical differences in the level of IL-10 in peripheral blood among the 3 groups (P ＞ 0.05).In the children with RSV infection,the expression of CD44 in peripheral blood was positively correlated with the level of IL-4 (r =0.798,P ＜ 0.05),the level of IL-12 was negatively correlated with the level of IL-4 (r =-0.186,P ＜0.05).Conclusion The expression of adhesion molecule CD44 may play a role in the RSV infection induced bronchiolitis,and the high expression of CD44 in children with RSV infection may increase the susceptibility to bronchiolitis.
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Objective To knock down angiopoietin-1 expression in human gastric cancer cell line BGC-823 and to observe its effect of reversing tumor invasion. Methods siRNA sequence fragments was designed to target angiopoietin-1 and transferred into human gastric cancer cell line BGC-823. RT-PCR was used to assess the transcription level of angio-poietin-1 mRNA, then western blot and immunofluorescence were used to examine the expression level of three invasion-as-sociated proteins include integrinβ1, CD44V6 and Ang-1. Cell adhesion ability was evaluated by cell adhesion assay and cell invation was determined by matrigel and transwell plastic dual-chamber culture system. Results Ang-1 mRNA was knocked down by siRNA showed by RT-PCR. The expression of integrinβ1, CD44V6 and Ang-1 were significantly lower than control group(P<0.05), so did the cellular adhesion and invasion abilities(P<0.05). Conclusion Knocking down angiopoietin-1 by siRNA can reverse invasion of human gastric cancer cell line BGC-823 and may provide new ideas and reference for gene therapy of gastric cancer in the future.
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PURPOSE: To analyze the immunohistochemical expression of the standard isoform of CD44 (CD44s) adhesion molecule in clear cell renal cell carcinoma (CCRCC) and its impact on clinical outcomes. MATERIALS AND METHODS: Ninety-nine consecutive patients treated surgically for RCC between 1992 and 2009 were selected. A single pathologist reviewed all cases to effect a uniform reclassification and determine the most representative tumor areas for construction of a tissue microarray. The same pathologist, who was blinded to the outcome of the cases, semi-quantitatively scored the staining intensity of CD44s in all specimens. The counting was done using the H-Score algorithm. RESULTS: Of the 99 immunostained RCC specimens, 57(57.7%) showed low expression, and 42(42.4%) showed high expression levels of CD44s. The expression of CD44s was directly associated with tumor size (p = 0.03), clinical stage (p = 0.02) and Fuhrman grade (p = 0.02). Disease specific survival (DSS) rates for patients whose specimens expressed low and high levels of CD44s was 88.1% and 67.5%, respectively (p = 0.009). Progression free survival (PFS) rates in patients with low and high expression of CD44s were 78.8% and 61.7%, respectively (p = 0.05). Classical features such as the presence of metastasis and clinical stage remained isolated predictors of survival. CONCLUSIONS: Immunohistochemical expression of CD44s was associated with important clinical variables such as stage and Fuhrman grade. However, it was not an independent predictor of survival. Therefore, we believe it has a limited role as a prognostic marker in patients with CCRCC.
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Adult , Aged , Female , Humans , Male , Middle Aged , /analysis , Carcinoma, Renal Cell/immunology , Biomarkers, Tumor/analysis , Carcinoma, Renal Cell/chemistry , Carcinoma, Renal Cell/pathology , Epidemiologic Methods , Immunohistochemistry , Prognosis , Sex Distribution , Time Factors , Tissue Array AnalysisABSTRACT
Objective To investigate the effects of different concentrations of sevoflurane on adhesion and expression of CD24 and CD44v6 in human lung cancer cell line A549.Methods Human lung cancer cell line A549 was obtained from Shanghai Cell Biology Medical Research Institute,Chinese Academy of Sciences and cultured in RPMI1640 culture medium containing 10％ fetal calf serum.The cells were inoculated in 24 well culture plate.After being cultured for 24 h,the cells were randomly divided into 4 groups:control group (group C) and 3 sevoflurane groups exposed to 1.7 ％,3.4 ％ and 5.1％ sevoflurane for 2,4 and 6 h respectively ( groups S1,S2,S3 ).The cells were cultured for another 48 h.Cell adhesion rate was detected by adhesion test and the expression of CD24 and CD44v6 mRNA and protein was determined by RT-PCR and flow cytometry.Results Sevoflurane significantly inhibited the cell adhesion rate and down-regulated CD24 and CD44v6 expression in a concentration and duration of exposure-dependent manner.Conclusion Sevoflurane can inhibit cell adhesion through down-regulation of CD24 and CD44v6 expression.
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Objective: To explore the characteristics of leukocyte-derived microparticles (LMPs) in breast cancer patients and their clinical significance. Methods: The expressions of LMPs, CD44+ LMPs, CD24+ LMPs and CD44+ CD24- LMPs in peripheral blood of 44 breast cancer patients and 10 healthy volunteers were detected by using flow cytometry. The percentages of CD44+, CD24+ and CD44+ CD24- cells in monocytes (MNCs) of breast cancer pateints and healthy volunteers were further detected. The clinical relevance of LMPs, and the correlation between MNCs and LMPs were analyzed. Results: The percentages of LMPs, CD44+ LMPs and CD44+ CD24- LMPs in breast cancer patients were statistically higher than those in the healthy volunteers (P =0.021, P =0.011, and P =0.006, respectively). There was a statistical difference in the percentage of LMPs among the different tumor volunmes (P =0.018). Statistical difference was also obseved in the percentage of CD44+ LMPs among different Her-2 levels (P =0.020). The percentages of CD44+, CD24+ and CD44+ CD24- cells in MNCs in breast cancer patients were not statistically different from those in the healthy volunteers. There was no statistical correlation between MNCs and LMPs. Conclusion: The percentage of LMPs in breast cancer patients can be used to evaluate the tumor volume. The percentage of CD44+ LMPs was closely correlated with Her-2 level, and it can be used to evaluate the recurrence, metastasis and prognosis of breast cancer. Copyright© 2012 by TUMOR.
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Objective: To investigate the effect of cell adhesion molecule CD44v6 on biological behavior of bladder cancer cell line T24. Methods: The expressions of CD44v6 protein in bladder transitional epithelial cells and bladder cancer T24 cells were detected by immunocytochemistry. MTT assay was used to detect the growth inhibitory rate of T24 cells treated with different concentrations of anti-CD44v6 monoclonal antibodies for 24, 48 and 72 h, respectively. After T24 cells were treated with anti-CD44v6 monoclonal antibodies, the apoptosis was detected by flow cytometer, the expressions of apoptosis-related markers Bcl-2 and Bax were determined by immunohistochemistry, and the change of migration ability of T24 cells was detected by Transwell migration test. Results: CD44v6 was expressed in T24 cells, but rarely expressed in bladder transitional epithelial cells. MTT assay demonstrated that anti-CD44v6 monoclonal antibodies had inhibitory effects on the proliferation of T24 cells (P <0.01). The apoptosis rate of T24 cells was increased with an induction of anti-CD44v6 monoclonal antibodies (P <0.01). The expression level of Bax protein was up-regulated, while the expression level of Bcl-2 protein was down-regulated in T24 cells (P <0.01). The migration of T24 cells was inhibited after treatment with anti-CD44v6 monoclonal antibodies (P <0.05). Conclusion: Anti- CD44v6 monoclonal antibodies can inhibit the abilities of proliferation and migration of T24 cells, and also induce apoptosis. The mechanism may be related with the expression regulation of Bcl -2 and Bax genes. Copyright© 2012 by TUMOR.
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Objective To investigate the biological effect of epithelial-to-mesenchymal transition (EMT) under the treatment of transforming growth factor (TGF)-β1 on the human gastric cancer cell line SGC7901 in vitro,and to observe whether the TGF-β1 can generate the tumor initiating cells ability in SGC7901 or not.Methods SGC7901 cells were cultured with TGF-β1.The morphological change was observed.The effect on proliferation of SGC7901 cells was detected by CCK-8.The invasion assay was used to investigate the motility and the invasion ability of SGC7901 cells.Immuofluorescence was used to detect the expression of E-cadherin and N-cadherin.The mRNA and protein's expression levels of EMT-related factors and CD44 were analyzed by RT-PCR and Western blotting respectively.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of SGC7901 cells was inhibited,and the ability of motility and invasion of SGC7901 cells were greatly enhanced after being treated with TGF-β1.RT-PCR and Western blotting showed that the expression of Snail (P ＜ 0.05),N-cadherin (P ＜ 0.05) and CD44 (P ＜ 0.05) were significantly increased while the expression of E-cadherin was decreased (P ＜ 0.05).Conclusions TGF-β1 can generate the EMT.The CD44 expression was up-regulated.TGF-β1 can inhibit the proliferation and promote the motility and invasion ability of SGC7901 cells.