Msi1基因沉默对人肝癌HepG2细胞的影响  

Downregulation of Msil by RNAi in Effect of HepG2 Cell Line

作　　者：王燕[1] 章建国[1] 张曙[1] 靳钦[1] 卞婷婷[1] 

机构地区：[1]南通大学附属医院病理科,江苏226001

出　　处：《交通医学》2014年第2期115-118,共4页Medical Journal of Communications

摘　　要：目的:探讨siRNA沉默Msi1基因表达后,人肝癌HepG2细胞的增殖与凋亡的情况。方法:设计合成针对Msi1 mRNA序列的siRNA多条序列,分别转染人肝癌HepG2细胞。使用RT–PCR方法检测各组细胞Msi1基因表达情况;Western blot法检测survivin蛋白、caspase3蛋白表达情况;MTT法、平皿克隆实验检测HepG2细胞增殖情况;流式细胞术检测各组HepG2细胞凋亡情况。结果:(1)RT-PCR结果:转染后24 h后,序列a,b,c转染效率分别为65%,64%及62%。其抑制率分别是Msi1 siRNAa(68%)、Msi1 siRNAb(56%)、Msi1 siRNAc(35%)。(2)流式细胞术检测各组细胞凋亡:空白组、阴性组、脂质体组、Msi1 siRNAa组凋亡率分别为(4.14±0.26)%、(4.51±0.78)%、(4.19±1.21)%,(16.8±0.26)%,Msi1 siRNAa组与空白对照组比较差异有统计学意义(P<0.05)。(3)Msi1 siRNAa最能有效抑制肝癌HepG2细胞中Msi1表达,与空白对照组比较,Msi1 siRNAa组HepG2细胞生长、增殖速度明显减缓(P<0.05);survivin蛋白表达水平显著下调(P<0.05),而caspase3蛋白表达水平上调(P<0.05);Msi1 siRNAa组凋亡率增高(P<0.05)。结论:沉默Msi1可抑制人肝癌HepG2细胞的增殖,诱导其凋亡。Objective：So far, the role of Msi1 gene in human hepatoma HepG2 cells is not clear, this paper focuses on the influence of human hepatoma HepG2 cell proliferation and apoptosis caused by the Msi1 siRNA. Methods：Design and synthesis of Msi1-specific siRNA and transfect of HepG2 cells were conducted, and RT -PCR was used to detect the expression of cell Msi1 mRNA in each group. One group was selected to continue the follow-up experiments, Western blot was used to detect the expressions of survivin and caspase3 proteins, proliferation of HepG2 cells was detected by MTT as-say and plate cloning, the apoptosis of HepG2 cells in each group was detected by flow cytometry. Results：Msi1 siRNAa most effectively inhibited the expression of Msi1 in HepG2 cells ,compared with the control group, the proliferation rate of Msi1 siRNAa HepG2 cell slowed down significantly（P＆lt;0.05）, protein expression of survivin was significantly decreased （P＆lt;0.05）, while that of caspase3 increased（P＆lt;0.05）, The percentage of apoptotic cells with Msi1 siRNA cells was higher. Con-clusions：Downregulation of Msi1 resulted in significant inhibition of tumor growth and induced apoptosis.

关 键 词：原发性肝癌 Msi1基因 凋亡 增殖 HepG2细胞 逆转录聚合酶链反应 Western BLOT检测 流式细胞术 Msi1 Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Flow Cytometry(FCM) 

分 类 号：R735.7[医药卫生—肿瘤]