靶向survivin的siRNA联合5-Fu转染抑制肝细胞株HepG2增殖的研究

目的：研究靶向survivin的（小分子干扰RNA）siRNA和（氟尿嘧啶）5-FU联用对肝癌细胞HepG2的增殖抑制及凋亡的影响。方法：将HepG2细胞分为空白对照组、阴性对照组、5-FU处理组、siRNA转染组、5-FU＋siRNA转染组。转染采用脂质体法。RT-PCR法检测HepG2细胞survivinmRNA转录水平；MTT法检测靶向survivin的siRNA和5．FU对HepG2细胞增殖的抑制作用；流式细胞术检测HepG2细胞凋亡情况。结果：空白对照组、阴性对照组、5-FU处理组survivinmRNA表达无明显变化（P〉0．05），siRNA转染组、5-FU＋siRNA转染组survivinmRNA表达明显下降（F=280．326，q=4．72-7．34，P〈0．05）。5-FU＋siRNA转染组增殖抑制率为51．58％±1-35％，与其它各组相比抑制率明显增高（F=280．326，q=5．27-9．84，P〈0．05）。5-Fu＋siRNA组与其它各组相比细胞凋亡率明显增高（F=13568．68，q=110．47-327．16，P〈0．01）。结论：将靶向survivin的siRNA和5一Fu联合应用可以显著抑制肝癌细胞survivin基因表达，并协同抑制HepG2细胞增殖，共同发挥诱导细胞凋亡作用。

Study on the Antiproliferation Effect of Targeting of Survivin siRNA Transfection Combined with 5-FU on Liver Cell Line HepG2

Objective： To investigate the anti-proliferation effect of （small interference RNA） siRNA against survivin combined with（fluorouracil） 5-FU on human liver cancer cell line HepG2. Methods： HepG2 cells cultures were divided into five groups： blank con- trol group, negative control group, 5-FUgroup, siRNA-survivin group, siRNA-survivin＋5-FU group. Lipofectamine TM 2000 was used to transfect HepG2 cell. The expression of survivin mRNA was detected by RT-PCR. Inhibition rate of each group on HepG2 was assayed by MTT method, and apoptosis was analyzed by flow cytometry respectively. Results： The experiment of RT-PCR showed that the ex- pression of survivin mRNA in siRNA-survivin group and siRNA-survivin＋5-FU group were lower than that in other groups（P〈0.05）, but its levels in blank control group, negative control group and 5-FU group had no significant change （P〉0.05）. The MTT assay indicated that the antiproliferation rate of siRNA-survivin＋5-FU group was 51.38%＋ 1.35. The antiproliferation rate of this group was higher than that in any other groups（P〈0.05）. Flow cytometry results indicated that the apoptosis rate of siRNA-survivin＋5-FU group was also higher than any other groups （P〈0.05）. Conclusion： siRNA against survivin combined with 5-FU can specifically suppress its expression, inhibit the proliferation of HepG2 and induce apoptosis in common.