慢病毒介导的膜孔相互作用分子1基因敲减对SMMC7721细胞凋亡的影响  

Effects of lentivirus-mediated stromal interaction molecular 1 gene knock-down on apoptosis of SMMC7721 cells

作　　者：王弼[1] 陈燕凌[1] 张吉成[1] 蔡欣然[1] 

机构地区：[1]福建医科大学附属协和医院肝胆外科,福州350001

出　　处：《中华实验外科杂志》2012年第7期1309-1311,共3页Chinese Journal of Experimental Surgery

基　　金：福建省科技计划重点资助项目（2008Y0043）

摘　　要：目的观察膜孔相互作用分子1（Stiml）基因表达下调对肝细胞癌（HCC）细胞凋亡的作用。方法设计和合成Stiml基因干扰序列，构建慢病毒介导的RNA干扰载体，利用第3代慢病毒包装系统进行病毒制备；干扰慢病毒以2倍感染复数（MOI）感染SMMC7721细胞，定量聚合酶链反应（PCR）和免疫印迹法（WB）检测Stiml基因表达；采用碘化丙锭（PI）和膜联蛋白V（AnnexinV）双染法检测SMMC7721细胞的凋亡变化。结果测序结果表明干扰序列连入干扰载体；定量PCR和Westernblot法检测结果显示干扰慢病毒感染可以抑制Stiml基因75％以上的表达；流式细胞仪检测结果表明在Stiml基因被敲减后，凋亡细胞的比例从8％上升到26％。结论Stiml基因的敲减促进SMMC7721细胞的凋亡。Objective To observe the effect of stromal interaction molecular 1 （Stiml） down-reg- ulation on apoptosis of hepatocellular carcinoma （HCC） cells. Methods The siRNA to Stiml was de- signed and synthesized, inserted to lentiviral vector. The third generation envelope system was used to produce lentiviral particles. Lentivirus infection of SMMC7721 was performed at MOI = 2. The expression of Stiml was detected by using quantitative polymerase chain reaction （PCR） and Western blotting. Annexin V and proliferation index （PI） staining was used to evaluate apoptosis of SMMC7721 cells. Results Se- quencing confirmed Stiml siRNA fragment was linked into interference vector. The expression of Stiml in infected cells was knocked down by above 75% detected by quantitative PCR and Western blotting. FACS showed that SMMC7721 apoptosis rate was increased with knockdown of Stiml. Conclusion Down-regula- tion of Stiml promotes apoptosis of SMMC7721 cells.

关 键 词：癌 肝细胞 膜孔相互作用分子1 RNA干扰 脱噬作用 

分 类 号：R735[医药卫生—肿瘤]