蛋白激酶C-α反义核酸对人肝癌细胞HepG2体外增殖及凋亡的影响

Effects of antisense oligonucleotides of PKC-α on proliferation and apoptosis of HepG2 in vitro

目的 探讨蛋白激酶 (PK )C α反义寡核苷酸 (asODN)对人肝癌细胞HepG2体外增殖及凋亡的影响。方法 用RPMI 164 0体外培养和传代HepG2细胞 ,以脂质体 (LP)介导不同浓度PKC αasODN转染人肝癌细胞HepG2 ,四甲基偶氮唑蓝 (MTT)比色法检测其生长指数 (GI) ,软琼脂克隆形成法检测其克隆形成率 ,流式细胞术 (FCM )检测其凋亡率。结果 0 .10～ 1.0 0 μmol/L的PKC αasODN能显著降低HepG2细胞GI(P <0 .0 5 ) ;0 .0 5～ 1.0 0 μmol/L的PKC αasODN能显著降低其软琼脂克隆形成率 (P <0 .0 1) ,两者均具有量效依赖关系。 0 .5 0～ 1.0 0 μmol/L的PKC αa sODN可使HepG2细胞发生显著凋亡 (P <0 .0 1)。结论 通过PKC αasODN阻断人肝癌细胞HepG2信号转导分子PKC α ,可有效抑制HepG2体外生长增殖 。

Objective To investigate the effects of antisense oligonucleotides of PKC α on proliferation and apoptosis of human hepatoma cell line HepG2 in vitro.Methods The human liver cancer cell line HepG2 was cultured in RPMI?1640 medium and subcultured in vitro.Different concentrations of PKC α antisense oligonucleotides (asODN) and a random sequence as a control were transfected into HepG2 cells by lipofectin (LP).The cell growth index (GI) and the clone formation rate of HepG2 were detected by MTT colorimetric assay and soft agar assay,respectively.The apoptpsis rate of HepG2 treated with PKC α asODN was assayed by flow cytometry(FCM).Results The GI of HepG2 transfected by PKC α asODN with concentrations ranging from 0.10 μmol/L to 1.00 μmol/L were lower significantly than that of control groups ( P <0.05).The clone formation rates of HepG2 transfected by PKC α asODN from 0.05 μmol/L to 1.00 μmol/L were lower very significantly than that of control groups ( P <0.01),and there was a dose dependent relationship among them.The apoptpsis rates of HepG2 treated with PKC α asODN from 0.50 μmol/L to 1.00 μmol/L were significantly higher than that of control group.Conclution PKC α asODN could inhibit the growth and proliferation of HepG2 and induce its apoptpsis through blocking the cell signal transduction related to PKC in vitro.