| RNA干扰沉默CDKL1基因对人胶质瘤U251细胞增殖和凋亡的影响 |
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| 引用本文： | 王佳,王杰军,夏士安,陈音,吴国华,车莉萍.RNA干扰沉默CDKL1基因对人胶质瘤U251细胞增殖和凋亡的影响[J].上海医学,2011,34(5). |
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| 作者姓名： | 王佳 王杰军 夏士安 陈音 吴国华 车莉萍 |
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| 作者单位： | 1. 上海交通大学医学院附属新华医院肿瘤科,200092
2. 第二军医大学附属长征医院肿瘤科 |
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| 摘 要： | 目的 利用RNA干扰(RNAi)在人胶质瘤U251细胞中沉默CDKL1(cyclin-dependent kinaselike 1)基因的表达,探讨其对肿瘤细胞增殖和凋亡的影响.方法 构建针对CDKL1基因的短发卡(shRNA)慢病毒表达载体,包装成病毒颗粒并感染人胶质瘤U251细胞,采用实时定量聚合酶链反应和Western印迹法验证CDKL1基因的mRNA和蛋白的表达,确定其抑制效率.采用甲基噻唑基四唑(MTT)比色法和应用Annexin V染色流式细胞仪检测细胞增殖和凋亡情况,观察沉默CDKL1基因对U251细胞增殖与凋亡表型的影响.结果 CDKL1-shRNA能有效抑制U251细胞中CDKL1基因的mRNA和蛋白的表达.CDKL1基因沉默后U251各时相的光密度值较感染无义序列的细胞均有降低趋势,第5天时差异有统计学意义(P＜0.05).同时,抑制CDKL1的U251细胞凋亡比例从无义小干扰RNA(siRNA)序列对照组的(9.80±0.21)%上升至(19.26±0.33)%,差异有统计学意义(P＜0.05).结论 采用RNAi方法 能有效沉默CDKL1基因的表达,同时显著降低U251细胞的增殖能力,促进肿瘤细胞凋亡,表明CDKL1基因可能具有影响胶质瘤发生、发展的作用.
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| 关 键 词： | RNA干扰 胶质瘤 增殖 凋亡 |
Effect of silencing cyclin-dependent kinase-like 1 gene by RNA interference on proliferation and apoptosis in human U251 glioma cells |
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| WANG Jia,WANG Jiejun,XIA Shian,CHEN Yin,WU Guohua,CHE Liping.Effect of silencing cyclin-dependent kinase-like 1 gene by RNA interference on proliferation and apoptosis in human U251 glioma cells[J].Shanghai Medical Journal,2011,34(5). |
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| Authors: | WANG Jia WANG Jiejun XIA Shian CHEN Yin WU Guohua CHE Liping |
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| Institution: | WANG Jia,WANG Jiejun,XIA Shian,CHEN Yin,WU Guohua,CHE Liping. Department of Oncology,Xinhua Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200092,China |
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| Abstract: | Objective To investigate the effect of silencing cyclin-dependent kinase-like 1(CDKL1) gene by RNA interfering on proliferation and apoptosis in human U251 glioma cells.Methods Small interference RNA(siRNA) targeting CDKL1 gene was cloned into lentivirus vector.Then shRNA lentivirus particles were packaged and infected into U251 cells.Real-time polymerase chain reaction(PCR) and Western blotting analysis were performed to investigate the expression of CDKL1 at mRNA and protein levels.The cell proliferation ... |
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| Keywords: | CDKL1 |
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