中国病理生理杂志2012，Vol.28Issue(7)：1241-1246,6.DOI:10.3969/j.issn.1000-4718.2012.07.017
siRNA-survivin与GRIM-19共表达质粒的构建及其对前列腺癌DU145细胞生长的抑制作用
Construction of siRNA-survivin and GRIM-19 coexpression plasmid and its growth inhibitory effect on prostatic cancer DU145 cells
摘要
Abstract
ADM: To construct the recombinant plasmid that expresses siRNA — survivin and GRIM — 19 simultaneously , and to identify the validity of the recombinant plasmid and observe its effect on expression of survivin and GRIM - 19 and proliferation ability of prostate cancer DU145 cells. METHODS: The recombinant plasmid coexpressing siRNA — survivin and GRIM — 19 was constructed using gene cloning technique. The prostatic cancer DU145 cells were transfect-ed with the coexpression plasmid and control plasmids. Survivin and GRIM — 19 mRNA expression was detected by semi — quantitative RT - PCR. The proliferation ability affected by coexpression plasmid was measured by MTT assay. RESULTS : The coexpression plasmid pGRIM — 19 — si — survivin was successfully constructed according to DNA recombinant technique and identified through restriction enzyme digestion and plasmid sequencing. Compared with the mock, survivin mRNA expression levels were 0. 55 ±0. 05,0. 62 ±0. 08 and 0. 35 ±0. 05 in psi - survivin, pGRIM - 19 and pGRIM - 19 - si - survivin groups, respectively. Compared with psi - survivin and pGRIM - 19, pGRIM - 19 - si - survivin inhibited survivin mRNA expression markedly ( P < 0. 05 ), while the expression levels of GRIM — 19 mRNA were 1. 93 ±0. 14, 2. 57 ±0. 20 and 4. 12 ±0. 21 in psi - survivin, pGRIM - 19 and pGRIM - 19 - si - survivin groups, respectively ( P < 0. 01 ). Compared with pGRIM — 19 group, pGRIM — 19 — si — survivin enhanced GRIM — 19 mRNA expression more obviously ( P < 0. 05 ). After transfection for 48 h, the proliferation rates were 58. 0% ± 7. 2% , 62. 1 % ± 6. 1 % and 50. 2% ± 4. 8% in the 3 experiment groups compared with the mock ( P < 0. 05 ). After transfection for 72 h, the proliferation rate were 43. 4% ±4. 3% , 51. 3% ±6. 7% and 26. 8% ±7. 1% in experiment groups compared with the mock ( P <0. 05 ). Compared with psi — survivin and pGRIM — 19, pGRIM — 19 — si — survivin significantly inhibited the cell growth ( P < 0. 05 ). CONCLUSION: Transfection of coexpression plasmid pGRIM — 19 — si — survivin dramatically changes the mRNA expression of survivin and GRIM - 19 and inhibits the cell growth.关键词
前列腺肿瘤/存活蛋白/GRIM-19Key words
Prostatic neoplasms/ Survivin/ GRIM - 19分类
医药卫生引用本文复制引用
沈维高,赵丽晶,盖晓东,芦丽莉,葛贺,陈立松,刘艳波,赵雪俭..siRNA-survivin与GRIM-19共表达质粒的构建及其对前列腺癌DU145细胞生长的抑制作用[J].中国病理生理杂志,2012,28(7):1241-1246,6.基金项目
国际科技合作重点资助项目(No.2004DFB02000) （No.2004DFB02000）
吉林省教育厅自然科学研究计划项目(No.2011146) （No.2011146）
吉林市科技计划项目(No.201032235) （No.201032235）
