ARF-BP1基因对肝癌 HepG2细胞凋亡的影响及其机制探讨

摘要

Objective To investigate the effect of ARF-BP1 gene on apoptosis of hepatoma HepG2 cells, and to ex-plore its mechanism.Methods HepG2 cells in logarithmic growth phase were randomly divided into 4 groups.Transfec-tion group was transfected translently by 100 nmol/L ARF-BP1 siRNA with LipofectamineTM 2000;liposome control group was only added liposomes; negative control group was transfected by negative siRNA fragments; blank control group was added the same amount of medium only, without siRNA fragments and liposomes.The flow cytometry was utilized to detect cells apoptosis rate of 4 groups; RT-PCR was used to detect the relative expression of p53, Mcl-1 mRNA in transfection group and blank control group.Results The apoptosis rate of transfection group, liposome control group, negative control group and blank control group were 27.90%±1.40%, 3.33%±0.66%, 3.05%±0.73% and 1.64%±0.12%, re-spectively;transfection group compared with other groups, all P<0.01; negative control group and liposome group com-pared with the control group, both P<0.05.72 h after transfection, p53, Mcl-1 mRNA relative expression in transfection group were 0.29 ±0.08 and 0.23 ±0.04, significantly lower than those in the blank control group ( P<0.01 and P<0.05).Conclusion Reduced ARF-BP1 gene expression in HepG2 cells can promote tumor cell apoptosis, which maybe correlate with inhibition of p53, Mcl-1 expression.%目的：观察ARF-BP1基因对肝癌HepG2细胞凋亡的影响，并探讨其机制。方法取对数生长期的HepG2细胞，随机分为4组。转染组采用脂质体LipofectamineTM2000包裹技术将100 nmol/L ARF-BP1 siRNA瞬时转染HepG2细胞；脂质体对照组只加脂质体，不加siRNA片段；阴性对照组转染阴性siRNA片段；空白对照组不加siRNA片段及脂质体，仅加等量培养基。采用流式细胞术检测各组细胞凋亡率，RT-PCR法检测转染组和空白对照组p53、Mcl-1 mRNA相对表达。结果转染组、脂质体对照组、阴性对照组和空白对照组的细胞凋亡率分别为27．90％±1．40％、3．33％±0．66％、3．05％±0．73％和1．64％±0．12％；转染组与其他组比较，P均＜0．01；阴性对照组、脂质体对照组与空白对照组比较，P均＜0．05。转染72 h后，转染组p53、Mcl-1 mRNA相对表达分别为0．29±0．08和0．23±0．04，均低于空白对照组的0．63±0．07和0．34±0．02，P＜0．01、0．05。结论降低肝癌HepG2细胞ARF-BP1基因表达会促进肿瘤细胞凋亡，可能与其抑制p53、Mcl-1基因表达有关。