RNA干扰抑制RPS12基因表达对胃癌细胞凋亡的影响及分子机制探讨  

The Effects of Silencing the RPS12 Gene by RNAi on Apoptosis of Gastric Cancer Cells and the Possible Molecular Mechanism

作　　者：闫杨[1] 付浩[1] 滑君[1] 张瑞秀[1] 陈丹琦[1] 孙开来[1] 孙秀菊[1] 

机构地区：[1]中国医科大学医学遗传学教研室,沈阳市110001

出　　处：《中国肿瘤临床》2008年第24期1415-1418,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金(编号:30570848);辽宁省自然科学基金资助(编号:20032075)~~

摘　　要：目的:探讨RPS12基因特异的RNA干扰对胃癌BGC823细胞凋亡的影响及机制。方法:设计并构建RPS12特异性shRNA表达载体(RPS12-shRNA),以Lipofectamine2000介导将RPS12特异性shRNA及Scram-ble-shRNA表达载体分别转染胃癌细胞。RT-PCR方法检测BGC823细胞中RPS12基因及凋亡相关基因Bcl-2和Bax mRNA的表达,流式细胞术(flow cytometry,FCM)分析RNA干扰对细胞凋亡的影响。结果:RT-PCR结果显示,RPS12-shRNA表达载体转染第3、5、7天后,RPS12基因表达低于对照组,其中第5天下调最明显。RPS12特异性RNA干扰第5天、第7天后,Bcl-2 mRNA表达明显下调,BaxmRNA表达无明显改变。流式细胞术分析结果显示,RNA干扰RPS12基因第7天后,胃癌细胞凋亡率(17.84%)明显高于对照组(1.89%)。结论:RNA干扰RPS12基因促进胃癌细胞凋亡作用可能是其作为上游基因通过下调Bcl-2基因表达而实现的。Objective： To observe the influence of RPS12-specific RNA interference on the apoptosis of BGC823 gastric cancer cells and to determine a possible mechanism. Methods： The RPS12-specific shRNA expression vector was designed, constructed and transfected into gastric cancer cells using lipofectamine 2000. Scramble-shRNA was used as the control. The expression of RPS12 and apoptosis-associated genes Bcl-2 and Bax was examined by RT-PCR. Apoptosis of gastric cancer cells was detected by flow cytometry. Results： RT-PCR showed that RPS12 mRNA expression was markedly downregulated on day 3, 5, and 7 after transfection of the RPS12-shRNA expression vector, especially on day 5. The expression of Bcl-2 mRNA was significantly downregulated on day 5 and 7 after transfection of the RPS12-shRNA vector. No changes were found in Bax mRNA expression. On day 7 after transfection, the apoptosis rate of the transfected cells （17.84%） was higher than that of the control cells （1.89%）. Conclusion： Suppression of the RPS12 gene with RNAi can increase apoptosis of gastric cancer cells, possibly by downregulating the upstream expression of Bcl-2.

关 键 词：RNA干扰 RPS12 胃癌细胞 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]