CAJ | 학술논문

目的观察膜孔相互作用分子1(Stim1)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Stim1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Stim1基因表达；采用碘化丙锭(PI)和膜联蛋白V( Annexin V)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West-em blot法检测结果显示干扰慢病毒感染可以抑制Stim1基因75％以上的表达；流式细胞仪检测结果表明在Stim1基因被敲减后,凋亡细胞的比例从8％上升到26％.结论Stim1基因的敲减促进SMMC7721细胞的凋亡.
목적 관찰막공상호작용분자1(Stim1)기인표체하조대간세포암(HCC)세포조망적작용.방법 설계화합성Stim1기인간우서렬,구건만병독개도적RNA간우재체,이용제3대만병독포장계통진행병독제비；간우만병독이2배감염복수( MOI)감염SMMC7721세포,정량취합매련반응(PCR)화면역인적법(WB)검측Stim1기인표체；채용전화병정(PI)화막련단백V( Annexin V)쌍염법검측SMMC7721세포적조망변화. 결과 측서결과표명간우서렬련입간우재체；정량PCR화West-em blot법검측결과현시간우만병독감염가이억제Stim1기인75％이상적표체；류식세포의검측결과표명재Stim1기인피고감후,조망세포적비례종8％상승도26％.결론Stim1기인적고감촉진SMMC7721세포적조망.
Objective To observe the effect of stromal interaction molecular 1 ( Stim1 ) down-regulation on apoptosis of hepatocellular carcinoma (HCC) cells.Methods The siRNA to Stiml was designed 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 Sequencing 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-regulation of Stiml promotes apoptosis of SMMC7721 cells.