摘要
目的:探讨survivin-siRNA对前列腺癌细胞株PC-3M的增殖抑制作用。方法:构建两个针对survivin siRNA表达载体,与脂质体和空质粒两个对照组一起分别转染前列腺癌PC-3M细胞后,利用半定量RT-PCR和Western blotting分别检测细胞survivin mRNA及蛋白表达水平,MTT法测定细胞的增殖能力,流式细胞术检测细胞周期和凋亡。结果:转染72h后,两个实验组细胞的survivin mRNA水平是脂质体组的48%±6%(n=3)和30%±5%(n=3),蛋白表达水平是脂质体组的38%±4%(n=3)和36%±4%(n=3)。MTT检测实验组细胞的生长速度分别是脂质体组的44.20%±2.08%(n=3)和39.20%±1.93%(n=3),流式细胞术发现实验组G1期细胞比例明显高于两个对照组,而G2期和S期细胞比例减少,细胞出现凋亡。结论:利用survivin-siRNA可明显抑制前列腺癌PC-3M细胞的增殖,细胞受阻于G1期,诱导细胞凋亡,为进一步进行动物体内研究奠定了基础。
AIM： To study the inhibitory effects of survivin siRNAs on the growth of PC -3M cells. METHODS： Two pairs of DNA template coding siRNA against survivin were synthesized to construct two recombinant plasmids, psi -surl and psi -sur2. The two recombinants and the two controls, lipofeetin and vacant plasmid were transfected into PC-3M cells. The expressions of survivin mRNA and protein were detected respectively by RT- PCR and Western blotting. Proliferation abilities were measured by MTT, and the cell cycle and apoptosis were assayed by FCM. RESULTS： After 72 h of transfection, the level of cell survivin mRNA in the two siRNA groups was 48% ±6% （ n =3） and 30% ±5% （n =3） of that in lipofectin group, and expression of survivin protein were 38% ±4% （n=3） and 36% ±4% （n = 3） respectively of that in lipofectin control. The proliferation rate of cells in pSi - surl and psi - sur2 groups was also in- hibited according to MTT, about 44. 20% ＋ 2.08% （ n = 3 ） and 39. 20% ± 1.93% （ n = 3 ） of that in lipofeetin group. Cell numbers of G1 phase in two siRNA groups were significantly higher than that in two controls, while cells of G2 phase and S phase were much lower. Cell apoptosis was found in both siRNA groups. CONCLUSION： The two survivin siRNA significantly inhibit the expression of survivin in mRNA and protein levels, arrest the cell cycle in G1 phase, and suppress the growth of PC-3M cells and induce apoptosis in vitro.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第10期1873-1876,共4页
Chinese Journal of Pathophysiology
基金
国际科技合作重点项目(No.2004DFB02000)
