干扰RNA沉默tpd52基因对胶质瘤细胞增殖及凋亡的影响

摘要

Obiective To explored the effect of silencing tumor protein D52(tpd52) gene by RNA interference on proliferation and apoptosis in glioma celIs.Methods The lentivirus, expressing shRNA-tpd52(the nucleotide sequence of inhibiting tpd52) and shRNA-NC(the nucleotide sequence of negative control) were infected into human U87 glioma cells in vitro.In 48 h after transfection, the counts of expression GFP cells was measured by fluorescence microscope, and the expression level of TPD52 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot analysis.MTT assay was performed to evaluated cells proliferation.Cell apoptosis and cycle were detected by the flow cytometry with Annexin V and PI.Results The transfection rate of U87 cells was more than 90%.In contrast with the shRNA-NC group and the blank control group, the expression of TPD52 mRNA and protein of shRNA-tpd52 group were decreased significantly(P90%,与shRNA-NC组及空白对照组相比较,shRNA-tpd52组细胞TPD52 mRNA及蛋白表达量明显减少(P<0.01),而且细胞增殖能力明显下降,差异有统计学意义(P<0.05);细胞周期被阻滞在G0/G1期、细胞凋亡比例明显增加(P<0.05).结论 沉默胶质瘤细胞中tpd52基因表达后,可以有效抑制细胞增殖,阻滞细胞周期,促进细胞凋亡.