| ShRNA靶向干扰EGFR抑制结直肠癌细胞增殖的机制 |
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| 引用本文： | 闵寒,陈志荣,张舒羽,周俊东,胡群超,孟星君,吴锦昌.ShRNA靶向干扰EGFR抑制结直肠癌细胞增殖的机制[J].世界华人消化杂志,2011(33). |
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| 作者姓名： | 闵寒 陈志荣 张舒羽 周俊东 胡群超 孟星君 吴锦昌 |
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| 作者单位： | 南京医科大学附属苏州医院;苏州市立医院东区消化科;苏州大学放射医学与公共卫生学院;苏州市立医院东区肿瘤放疗科; |
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| 基金项目： | 国家自然科学基金资助项目,No.81102078; 苏州市科教兴卫青年科技基金资助项目,No.SWKQ1026; 苏州市科技发展指导基金资助项目,No.SZD09132 |
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| 摘 要： | 目的:从细胞周期的角度探讨RNA干扰抑制表皮生长因子受体(epidermal growth factor receptor,EGFR)表达对结直肠癌细胞增殖影响的机制.方法:以人结直肠癌细胞HCT-15为研究对象,构建EGFR-短发夹RNA(short hairpin RNA,shRNA)载体,采用脂质体转染的方法转染细胞,G418筛选稳定细胞株以获得稳定的转染细胞模型,分为4组:转染shRNA-NC,转染shRNA-1组,转染shRNA-2组,转染shRNA-3组;应用半定量RT-PCR和流式细胞术检测转染细胞模型的mRNA和蛋白表达水平;应用平板克隆实验检测转染后细胞增殖能力;应用流式细胞术检测转染后细胞周期的变化.结果:正确构建了shRNA质粒表达载体,成功转染;转染shRNA-1、2、3组较转染shRNA-NC组mRNA、蛋白表达水平均有下降,以转染shRNA-1和2组的抑制效应好(40.2%±3.2%,52.8%±11.3%);转染shRNA-1、2两组较对照组增殖能力下降,G0/G1期细胞增多(63.69±2.75%,60.10±2.00%vs51.08±3.42%)、S期细胞减少(28.20%±...
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| 关 键 词： | RNA干扰 表皮生长因子受体 细胞周期 结直肠癌 |
ShRNA-mediated silencing of the epidermal growth factor receptor gene inhibits proliferation of human HCT-15 colorectal cancer cells |
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| Han Min,Zhi-Rong Chen.ShRNA-mediated silencing of the epidermal growth factor receptor gene inhibits proliferation of human HCT-15 colorectal cancer cells[J].World Chinese Journal of Digestology,2011(33). |
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| Authors: | Han Min Zhi-Rong Chen |
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| Institution: | Han Min,Zhi-Rong Chen,Department of Gastroenterology,Suzhou Municipal Hospital (East Branch) Affi liated to Nanjing Medical University,Suzhou 215001,Jiangsu Province,China Shu-Yu Zhang,School of Radiation Medicine and Public Health,Medical College of Soochow University,Suzhou 215000,China Jun-Dong Zhou,Qun-Chao Hu,Xing-Jun Meng,Jin-Chang Wu,Department of Radiation Oncology,Suzhou Municipal Hospital (East Branch) Aff iliated to Nanjing Medical University,Jiang... |
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| Abstract: | AIM: To examine the effect of short hairpin RNA(shRNA)-mediated silencing of the epidermal growth factor receptor (EGFR) gene on the proliferation of human colorectal cancer cells and to explore the potential mechanisms involved. METHODS: Vectors containing shRNA targeting EGFR were constructed. HCT-15 cells were transfected with EGFR-shRNA-1, EGFR-shRNA-2, EGFR-shRNA-3 or shRNA-NC expression vectors, and stably transfected cell lines were screened. The mRNA level of EGFR was assessed by semi-quantitative R... |
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| Keywords: | RNAi Epidermal growth factor receptor Cell cycle Colorectal cancer |
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