【摘要】 目的探讨生存素(survivin)基因对肝癌Huh7细胞增殖、迁移和凋亡的影响。方法将肝癌细胞分为空白对照组(A组)、空白载体病毒组(B组)和ShRNA-survivin慢病毒组(C组),构建ShRNA-survivin慢病毒载体,通过RT-qPCR检测凋亡因子半胱氨酸天冬氨酸蛋白酶-9(Caspase-9)以及Caspase-3 mRNA的表达量;通过CCK-8实验检测肝癌Huh7细胞增殖能力;通过Transwell小室实验检测肝癌Huh7细胞迁移能力;通过AV-PI凋亡试剂盒检测肝癌Huh7细胞的凋亡情况。结果 A、B、C组survivin mRNA的相对表达量分别为1.000±0.000、1.079±0.011、0.421±0.097,A、B组与C组比较差异具有显著统计学意义(F=15.984,q=8.141、3.257,P<0.05),Caspase-3和Caspase-9的mRNA表达量与survivin的mRNA表达量趋势相同;A、B、C组中透过分子筛的细胞数比较差异有显著性(F=31.982,P<0.05);A、B组与C组比较差异均有显著意义(q=7.153、9.074,P<0.05);A、B、C组细胞的增殖率比较差异有显著意义(F=23.983,P<0.05);A、B组与C组比较差异均有显著意义(q=10.322、9.886,P<0.05);A、B、C组细胞的凋亡率比较差异有显著意义(F=30.009,P<0.05);A、B组与C组比较差异均有显著意义(q=9.531、8.001,P<0.05)。结论 ShRNA-survivin对肝癌Huh7细胞的增殖、迁移有抑制作用,对细胞凋亡具有促进作用,可以作为肝癌的潜在治疗靶点。更多还原

【Abstract】 Objective To explore the effects of survivin gene on the proliferation,migration,and apoptosis of hepatoma Huh7 cells. Methods Hepatoma cells were divided into blank control group(A),empty lentivirus vector group(B),and ShRNA-survivin lentivirus vector group(C).A ShRNA-survivin lentivirus vector was constructed.RT-qPCR was used to determine the mRNA expression of Caspase-9 and Caspase-3.CCK-8 kit,Transwell chamber,and AV-PI apoptosis kit were used to evaluate the proliferation,migration,and apoptosis of hepatoma Huh7 cells,respectively. Results There were significant differences between groups A,B,and C in the relative expression of survivin mRNA(1.000±0.000 vs 1.079±0.011 vs 0.421±0.097,F=15.984,q=8.141,3.257,P<0.05).The mRNA expression of Caspase-3 and Caspase-9 exhibited the same trends as that of survivin.There was a significant difference in the number of cells passing through the molecular sieve between the three groups(F=31.982,P<0.05),especially between group A/B and group C(q=7.153,9.074,P<0.05);there was a significant difference in the proliferation rate of cells between the three groups(F=23.983,P<0.05),especially between group A/B and group C(q=10.322,9.886,P<0.05);there was also a significant difference in the apoptosis rate between the three groups(F=30.009,P<0.05),especially between group A/B and group C(q=9.531,8.001,P<0.05). Conclusion The shRNA-survivin can inhibit the proliferation and migration and promote the apoptosis of hepatoma Huh7 cells.It can be used as a potential therapeutic target for hepatocellular carcinoma.更多还原