MMP-9-siRNA对乳腺癌细胞中MMP-9表达的影响

Effects of MMP-9-siRNA on MMP-9 Expression in MDA-MB-231 Cells

目的：探讨MMP-9小分子干扰RNA（MMP-9-siRNA）对乳腺癌MDA-MB-231细胞中MMP-9表达的影响.方法：化学合成针对MMP-9的小分子干扰RNA（MMP-9-siRNA）,并将其转染到乳腺癌MDA-MB-231细胞.本实验分为空白组（不转染任何siRNA）、对照组（转染Control-siRNA 50 nmol/L）和实验组（又分为3个亚组,分别转染10、50、100 nmol/L MMP-9-siRNA）3组.应用RT-PCR法及Western blot法分别在mRNA水平及蛋白水平检测并比较空白组、对照组和实验组中MMP-9表达及其差异.结果：RT-PCR结果显示,对照组MMP-9 mRNA表达率为99.2%±4.9%,与空白组比较差异无统计学意义（P＞0.05）;实验组10、50、100 nmol/L MMP-9-siRNA 3个亚组MDA-MB-231细胞MMP-9 mRNA表达率分别为80.3%±5.0%、65.2%±4.5%、55.9%±5.1%,与空白组比较差异有统计学意义（P＜0.05）.Western blot结果显示,对照组蛋白表达率为101.7%±3.1%,与空白组比较差异无统计学意义（P＞0.05）;实验组 10、50、100 nmol/L MMP-9-siRNA 3个亚组MDA-MB-231细胞MMP-9蛋白质表达率分别为77.6%±3.9%、62.7%±4.1%、55.0%±4.8%,与空白组比较差异有统计学意义（P＜0.05）.结论：构建的MMP-9-siRNA能够抑制MDA-MB-231细胞中MMP-9的表达,为基因水平治疗乳腺癌提供实验依据.

Objective：To investigate the effects of siRNA on MMP-9 expression in MDA-MB-231 cells. Methods： siRNA of targeting MMP-9 was designed and synthesized, then transfected into MDA-MB-231 cells. This study was divided into non - siRNA group, control group w^ich transfectd with Control- siRNA and exper imental group which transfected with MMP-9-siRNA. The experimental groups were transfected with 10, 50, 100 nmol/L MMP-9-siRNA respectively. MMP-9 mRNA expression was determined by RT- PCR and protein expression was detected by Western -blot analysis. Results：The results of RT -PCR showed, that MMP -9 mR- NA expression rate of control group was 99.2% ± 4.9%. There was no significance than normal group（ P 〉 0. 05 ）. The MMP -9 mRNA expression rate of experiment group in MDA-MB-231 ceils was decreased to 80.3% ±5.0%, 65.2% ±4.5% ,55.9% ±5.1% by 10,50,100mmol/L MMP -9 - siRNA respectively. There was significance than normal group （ P 〈 0.05 ）. The results of Western blot showed that protein expression rate of control group was 101.7%± 3.1%. There was no significance than normal group（ P 〉 0.05 ）. The MMP- 9 mRNA expression rate of experiment group in MDA-MB-231 cells was decreased to 77.6% ± 3.9% ; 62.7% ± 4.1% ,55.0% ± 4.8% by 10,50,100mmol/L MMP - 9 - siRNA respectively. There was significance than normal group（ P 〈 0.05 ）. Conclusions： MMP-9-siRNA can effectively down - regulate MMP-9 gene expression in MDA-MB-231 cell lines, which can provide a novel approach for therapeutic intervention to treat breast canc- er.