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机构地区:[1]上海交通大学医学院附属仁济医院临床干细胞研究中心,上海200127
出 处:《肿瘤》2014年第7期579-583,共5页Tumor
基 金:国家重大科学研究计划项目(编号:2012CB966800);国家自然科学基金资助项目(编号:81130038;81270627;81372189);上海市科学技术委员会科研计划项目(编号:12PJ1406100);上海市教育委员会科研创新项目(编号:13YZ030);上海市浦江人才计划资助项目(编号:D类17)
摘 要:目的 :探讨干细胞相关标志物和上皮-间质转化(epithelial-mesenchymal transition,EMT)相关基因在雄激素敏感和雄激素抵抗的前列腺癌细胞膜上的表达及前列腺癌干细胞中EMT相关基因的表达。方法 :应用FCM法检测前列腺癌雄激素敏感的LNCaP细胞和雄激素抵抗的DU145、PC3和C42B细胞膜上CD44、TRA-1-60、E-钙黏蛋白和N-钙黏蛋白的表达。实时荧光定量PCR法检测流式分选获得的TRA-1-60+和TRA-1-60-的DU145、PC3和C42B细胞中E-钙黏蛋白和N-钙黏蛋白mRNA的表达。结果 :雄激素抵抗的DU145、PC3和C42B细胞膜上CD44、TRA-1-60和N-钙黏蛋白的阳性表达率[(93.23±2.14)%、(85.54±1.52)%和(92.52±1.69)%,(0.95%±0.23)%、(0.59±0.31)%和(0.99±0.16)%,(2.23±0.53)%、(15.31±1.13)%和(19.33±1.58)%]高于雄激素敏感的LNCaP细胞[(0.11±0.03)%、(0.07±0.01)%和(0.11±0.02)%],而E-钙黏蛋白的阳性表达率[(55.23±2.57)%、(41.64±0.96)%和(59.72±1.36)%]低于雄激素敏感的LNCaP细胞[(96.31±1.98)%],差异均有统计学意义(P<0.01)。TRA-1-60+的前列腺癌细胞中N-钙黏蛋白mRNA的表达水平高于TRA-1-60-的前列腺癌细胞,而E-钙黏蛋白mRNA的表达水平低于TRA-1-60-的前列腺癌细胞,差异均有统计学意义(P<0.05)。结论 :雄激素抵抗的前列腺癌细胞中CD44、TRA-1-60和N-钙黏蛋白的阳性表达率较高,TRA-1-60+的前列腺癌干细胞中N-钙黏蛋白mRNA的表达水平较高。Objective: To investigate the membrane expressions of stem cell markers and epithelial-mesenchymal transition (EMT)-related genes in androgen-sensitive and androgen-resistant prostate cancer cell lines as well as the expressions of EMT-related genes in prostate cancer stem cells. Methods: The expressions of CD44, TRA-1-60, E-cadherin and N-cadheirn in androgen-sensitive prostate cancer LNCaP cells and androgen-resistant DU145, PC3 and C42B cells were detected by flow cytometry. The expressions of E-cadherin and N-cadheirn mRNAs in DU145, PC3 and C42B cells with TRA-1-60+ or TRA-1-60- were detected by real-time fluorescence quantitative-PCR. Results: The positive expression rates of CD44, TRA-1-60 and N-cadherin in androgen-resistant DU145, PC3 and C42B cells [(93.23±2.14)%, (85.54±1.52)%, (92.52±1.69)%; (0.95%±0.23)%, (0.59±0.31)%, (0.99±0.16)%; (2.23±0.53)%, (15.31±1.13)%, (19.33±1.58)%] were higher than those in androgen-sensitive LNCaP cells [(0.11±0.03)%, (0.07±0.01)%, (0.11±0.02)%], whereas the positive expression rates of E-cadherin [(55.23±2.57)%,(41.64±0.96)%, (59.72±1.36)%] were lower than those in androgen-sensitive LNCaP cells [(96.31±1.98)%] (all P 〈 0.01). The expression level of N-cadherin mRNA in TRA-1-60+ prostate cancer cells was higher than that in TRA-1-60- cells, whereas the expression level of E-cadherin mRNA was lower than that in TRA-1-60- cells (P 〈 0.05). Conclusion: The positive expression rates of CD44, TRA-1-60 and N-cadherin in androgen-resistant prostate cancer cells are higher, and the expression level of N-cadherin mRNA in TRA-1-60+ prostate cancer stem cells is higher.
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