靶向survivin的siRNA并氟尿嘧啶转染对HepG2增殖影响

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增殖的抑制作用及对凋亡的影响。方法将HepG2细胞分为空白对照组、阴性对照组、5-FU处理组、siRNA转染组及5-FU＋siRNA转染组。转染采用脂质体法。采用RT-PCR法分别检测HepG2细胞survivin mRNA转录水平;MTT法分别检测靶向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 siRNA转染组、5-FU＋siRNA转染组survivin mRNA表达较空白对照组、阴性对照组、5-FU处理组明显下降（F=326.78,q=5.78～7.12,P〈0.05）。5-FU＋siRNA转染组细胞增殖抑制率为51.58%±1.35%,与其他各组相比明显增高（F=1 293.24,q=15.31～82.26,P〈0.01）。5-FU＋siRNA转染组与其他各组相比细胞凋亡率明显增高（F=13 568.68,q=110.47～327.16,P〈0.01）。结论将靶向survivin的siRNA和5-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。

THE EFFECT OF TARGETING SURVIVIN sIRNA TRANSFECTION COMBINED WITH 5-FU ON LIVER CELL LINE HepG2

Objective To investigate the anti-proliferation effect of siRNA against survivin combined with fluorouracil（5-FU） on human liver cancer cell line HepG2. Methods HepG2 cells were divided into five groups as blank control,negative control,5-FU,siRNA-survivin,and siRNA-survivin＋5-FU.Lipofectamine TM 2000 was used to transfect HepG2 cells.The expression of survivin mRNA was detected by RT-PCR.The effect on cell proliferation and apoptosis were measured by MTT assay and flow cytometry,respectively.The apoptosis of HepG2 cells were detected by flow cytometry. Results The expression of survivin mRNA in siRNA-survivin group and siRNA-survivin＋5-FU group was lower than that in the other groups（F=326.78,q=5.78-7.12,P〈0.05）,The cytostasis rate in siRNA-survivin＋5-FU group was 51.38%±1.35%,with significantly higher than that in the other groups（F=1 293.24,q=15.31-82.26,P〈0.01）,and the apoptosis rate of this group was also higher than the other groups（F=13 568.68,q=110.47-327.16,P〈0.01）. Conclusion Combined use of siRNA against survivin with 5-FU can notably inhibit the expression of surviving gene in liver cancer cells,and suppress the proliferation of HepG2 cells,inducing apoptosis of cancer cells.