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| 靶向EGFR基因RNA干扰对人卵巢癌SKOV_3细胞增殖的抑制作用 | |
| 引用本文： | 郭启帅,黄曦,吴永忠,李少林.靶向EGFR基因RNA干扰对人卵巢癌SKOV_3细胞增殖的抑制作用[J].第三军医大学学报,2009,31(23):2338-2341. |
| 作者姓名： | 郭启帅 黄曦 吴永忠 李少林 |
| 作者单位： | 1. 重庆医科大学放射医学教研室,重庆,400016 2. 重庆医科大学附属第一医院血液科,重庆,400016 3. 重庆医科大学重庆市肿瘤,重庆,400016 |
| 摘 要： | 目的 利用RNA干扰技术下调表皮生长因子受体(epidermal growth factor receptor,EGFR)基因的表达,观察其对卵巢癌SKOV3细胞增殖的影响.方法 构建针对EGFR基因的siRNA真核表达载体,转染入卵巢癌SKOV_3细胞中,并筛选稳定表达siRNA的转化克隆,RT-PCR及Western blot分别检测EGFR mRNA和蛋白表达,流式细胞仪检测细胞周期和凋亡,克隆形成实验及MTY法检测细胞克隆形成率及细胞体外的增殖情况.结果 成功构建表达质粒pGenSil-HK、pGcnSil-EGFRI和pGenSil-EGFR2,并转染SKOV,细胞,通过G418筛选后获得稳定转染克隆,RT-PCR及Western blot分析表明转染pGenSil-EGFRI、pGenSil-EGFR2后的细胞EGFR mRNA和蛋白表达均受到了明显抑制,FGFR mRNA抑制率分别为41.87%和68.07%,蛋白表达抑制率分别为45.21%和70.25%.与未转染组和pGenSil-HK组相比,pGenSil-EGFR2组细胞凋亡比例显著增加,G_1期细胞增多,而S期细胞减少(P<0.05);转染pGenSil-HK组克隆形成率为(0.82±0.03),转染pGenSil-EGFR2组克隆形成率为(0.61±0.04),二者比较差异显著(P<0.05),MTT显示细胞培养36 h后,pGenSil.EGFR2组的细胞生长均显著低于未转染组和转染pGenSil-HK组(P<0.05).结论 靶向EGFR基因RNA干扰能够阻抑卵巢癌SKOV_3细胞中的EGFR表达,诱导细胞凋亡、调控细胞周期再分布、抑制细胞增殖. |
| 关 键 词： | RNA干扰 表皮生长因子受体 SKOV_3细胞 细胞周期 凋亡 |
RNA interference targeting EGFR on proliferation of human ovarian carcinoma cell line SKOV_3 |
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| GUO Qi-shuai,HUANG Xi,WU Yong-zhong,LI Shao-lin.RNA interference targeting EGFR on proliferation of human ovarian carcinoma cell line SKOV_3[J].Acta Academiae Medicinae Militaris Tertiae,2009,31(23):2338-2341. | |
| Authors: | GUO Qi-shuai HUANG Xi WU Yong-zhong LI Shao-lin |
| Institution: | GUO Qi-shuai1,HUANG Xi 2,WU Yong-zhong3,LI Shao-lin1 (1Department of Radiation Medicine,2Department of Hematology,First Affiliated Hospital,Chongqing Medical University,Chongqing 400016,3Cancer institute of Chongqing,Chongqing 400030,China) |
| Abstract: | Objective To explore the effect of down-regulating epidermal growth factor receptor (EGFR) expression by RNA interference on the proliferation of ovarian carcinoma SKOV3 cells. Methods The recombinant eukaryotic expression plasmids pGenSil-HK,pGenSil-EGFR1 and pGenSil-EGFR2 were constructed and transfected into SKOV3 cells respectively. The mRNA and protein expressions of EGFR in SKOV3 cells were detected by RT-PCR and Western blotting,respectively. The cell cycle and apoptosis were evaluated by flow cytome... |
| Keywords: | RNA interference epidermal growth factor receptor SKOV_3cell cell cycle apoptosis |
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