文献类型：期刊文献

中文题名：RUNX3小干扰RNA对SH-SY5Y细胞生长和药物敏感性的作用

英文题名：Effects of RUNX3 siRNA on the growth and drug sensitivity of SH-SY5Y cells

作者：张宏卫[1];刘丽华[2];王承忠[3]

机构：[1]绍兴文理学院医学院附属医院,浙江绍兴312000;[2]绍兴文理学院医学院附属医院分子诊断实验室,浙江绍兴312000;[3]中国科学院神经科学研究所,上海200031

年份：2008

卷号：24

期号：3

起止页码：504

中文期刊名：中国病理生理杂志

外文期刊名：Chinese Journal of Pathophysiology

收录：CSTPCD、、北大核心2004、CSCD2011_2012、北大核心、CSCD

语种：中文

中文关键词：神经母细胞瘤;RUNX3;RNA干扰;SH—SY5Y细胞

外文关键词：Neuroblastoma; RUNX3; RNA interference; SH -SY5Y cells

中文摘要：目的:探讨RUNX3基因对人神经母细胞瘤细胞生长和药物敏感性的调节作用。方法:构建RUNX3基因的小干扰RNA载体并将其转导入SH-SY5Y细胞,G418筛选后获得稳定转染的阳性克隆后,应用RT-PCR和Western blotitng进行鉴定;MTT法和流式细胞仪检测细胞转染前后生长速度和细胞周期的变化;MTT法和流式细胞仪检测细胞转染前后对阿霉素敏感性的变化;Western blotting检测细胞转染前后细胞周期相关蛋白cy-clin D1、CDK4、CDK6、p21、p27和凋亡相关蛋白Bcl-2、Bax以及耐药相关蛋白P-gp、MRP的表达变化。结果:成功构建了RUNX3的小干扰RNA载体并将其转染SH-SY5Y细胞;筛选到稳定的RUNX3低表达的神经母细胞瘤细胞模型;MTT和流式细胞仪检测结果显示,转染RUNX3小干扰RNA后的细胞的生长速度显著快于对照组(P<0.05),且G1期的细胞比率显著低于对照组(P<0.05);MTT法和流式细胞仪结果显示,转染RUNX3小干扰RNA后的细胞对化疗药物的敏感性降低,细胞内的阿霉素蓄积量显著减少(P<0.05);Western blotting显示,转染RUNX3小干扰RNA后的细胞中Bcl-2、P-gp和cyclin D1的表达明显增高,p21的表达明显降低。结论:下调RUNX3基因能促进神经母细胞瘤细胞生长,降低细胞对化疗药物的敏感性,提示RUNX3在神经母细胞瘤的发生和发展中可能扮演重要角色。

外文摘要：AIM： To investigate the effects of RUNX3 gene on the growth and drug sensitivity of SH - SYSY cells. METHODS： The siRNA plasmid of RUNX3 was constructed and transfected into SH -SY5Y cells. Stable transfectants were identified by RT - PCR and Western blotting. The growth curve, cell cycle distribution, drug sensitivity assay and accumulation of adriamycin in cells were detected by MTT assay and flow cytometry. The expressions of cyclin D1, CDK4, CDK6, p21, p27, Bcl - 2, Bax, P - gp and MRP were analyzed by Western blotting. RESULTS ： mU6pro - RUNX3 siR- NA was successfully constructed and transfected into SH -SYSY cells. Down -regulation of RUNX3 significantly promoted the cellular proliferation, inhibit the drug sensitivity and intracellular adriamycin accumulation of cells, compared with that in the controls （P 〈 0. 05）. The expressions of P - gp, Bcl - 2 and cyclin D1 in transfected cells were increased, while p21 decreased. CONCLUSION： RUNX3 might play important roles in the development of neuroblastoma.