论文部分内容阅读
目的探究沉默tpd52基因表达后对胶质瘤U87细胞增殖、周期及凋亡的影响。方法将携带着shRNA-tpd52(抑制tpd52的核苷酸序列)、shRNA-NC(阴性对照的核苷酸序列)的慢病毒体外转染胶质瘤细胞系U87。在转染48 h后荧光显微镜下观察表达GFP细胞数量,采用荧光定量PCR、Western blot分别检测TPD52 mRNA及蛋白表达量,MTT检测细胞增殖能力,流式细胞术检测细胞周期分布,Annexin V和PI双染流式细胞术检测细胞凋亡。结果 U87细胞转染率>90%,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。
Objective To investigate the effect of silencing tpd52 gene expression on the proliferation, cell cycle and apoptosis of glioma U87 cells. Methods Glioma cell line U87 was transfected in vitro with lentivirus carrying shRNA-tpd52 (nucleotide sequence of tpd52 inhibited), shRNA-NC (nucleotide sequence of negative control). 48 h after transfection, the number of GFP cells was observed under a fluorescence microscope. The expression of TPD52 mRNA and protein was detected by fluorescence quantitative PCR and Western blot respectively. The proliferation of cells was detected by MTT assay. Cell cycle distribution, Annexin V and PI Double staining flow cytometry to detect apoptosis. Results The transfection rate of U87 cells was> 90%. Compared with shRNA-NC group and blank control group, the expression of TPD52 mRNA and protein in shRNA-tpd52 group was significantly decreased (P <0.01), and the cell proliferation ability was significantly decreased (P <0.05). The cell cycle was arrested at G0 / G1 phase and the percentage of apoptosis was significantly increased (P <0.05). Conclusion The expression of tpd52 gene in glioma cells can effectively inhibit cell proliferation, arrest cell cycle and promote cell apoptosis.