摘要
目的合成凋亡抑制蛋白(hIAP-2)的si-RNA,观察其对hepG2细胞增殖、凋亡的影响。方法化学合成三条hIAP-2-siRNA干扰片段并用脂质体法转染hepG2细胞,同时设立脂质体对照。RT-PCR和western blotting检测hIAP-2-siRNA作用后hIAP-2mRNA和蛋白的表达,MTT检测细胞增殖,AV-PI流式细胞术检测细胞的凋亡。结果 RT-PCR及Western blootting检测显示3条siRNA均能有效抑制hIAP-2mRNA及蛋白表达(P<0.05),以siRNA1作用效果最显著(P<0.05)。MTT检测显示hIAP-2-siRNA80nmoL·L-1终浓度转染后的hepG2细胞的增殖受到显著抑制(P<0.01),其中以80nmoL·L-1转染72h时增殖性下降最明显(P<0.01)。流式细胞术检测结果显示hIAP-2-siRNA转染hepG2细胞后凋亡率增加(P<0.05)。结论 hIAP-2-siRNA可明显抑制该肿瘤细胞的增殖,促进hepG2肝癌细胞的凋亡。
Objective Observe the effects of siRNAs which are synthesized f^m human inhibitor of apoptosis protein 2 （hIAP-2） on the proliferation and apoptosis of HepG2 cells. Methods HepG2 cells were transfected through liposome mediated transfecfion method with 3 strands of hlAP-2-siRNA syn- thesized through chemical method, and the normal liposome were used as the control. The hIAP-2 mRNAand protein expression were tested using RT-PCR and Western blotting after hIAP-2-siRNAs taking ef- fects. The cell proliferation and apoptosis were detected by MTT and AV-PI flow cytometry, respectively. Results The results of RT-PCR and Western blotting showed that all of 3 pieces of siRNA can inhibit hIAP-2 mRNA and protein expression eff^tively （P〈0.05）, especially siRNA 1 （P〈0.05）. MTr showedthat the proliferation of HepG2 cells that were transfected by hlAP-2-siRNA of 80 nmoL.L^-1has been re- markably inhibited （P〈0.01）, and cell proliferation declined greatly after 7211 of transfection by hIAP-2- siRNA of 80 nmobL^-1 （P〈0.01）. The result of flow cytometry showed that the apoptosis rate of HepG2 cells tmnsfected by hIAP-2-siRNA raised （P〈0.05）. Conclusion Obviously, hlAP-2-siRNA can inhibit the orolifemtion of this tumor cells, and promote HepG2 liver cancer cells apoptosis.
出处
《肿瘤药学》
CAS
2011年第5期426-430,共5页
Anti-Tumor Pharmacy
