下调RNF138基因对胶质瘤细胞株U251增殖、凋亡及细胞周期影响的研究

目的探讨RNA干扰下调胶质瘤细胞株高表达基因RNF138对胶质瘤细胞株U251体外增殖、凋亡以及细胞周期的影响。方法构建下调RNF138基因的siRNA,通过慢病毒转染导入胶质瘤细胞株U251,设立阴性干扰对照组及空白对照组,荧光显微镜观察转染效率;实时荧光定量PCR检测敲减效率;运用Cellomics仪器连续检测U251体外增殖情况;流式细胞仪检测U251凋亡及细胞周期变化情况。结果慢病毒载体高效、稳定转染U251细胞,靶向RNF138的siRNA有效抑制RNF138基因表达,使RNF138mRNA表达减少60%,U251体外增殖明显减缓,凋亡明显增加,细胞阻滞在G2/M期。结论 RNA干扰抑制RNF138基因表达可以明显抑制胶质瘤细胞株U251体外增殖,促进其凋亡,影响细胞周期,阻滞细胞分裂。

Influence on the glioma cell line U251's proliferation, apoptosis and cycle after silencing RNF138 gene through RNA interference

Objective To explore the influence on the glioma cell line U251 ＇ s proliferation , apoptosis and cell cycle after silencing the high expressed gene RNF138 by RNA interference. Methods Firstly we constructed the RNF138-siRNA Lentivirus vector and infected the glioma cell line U251, the negative control group and blank control group were included. The transfected efficiency was observed by fluorescence microscope and the knockdown efficiency was detected by real time quantitative PCR. The effects of RNF138-konckdown on cell proliferation were assessed by Cellomics and cell apoptosis and cell cycle were assessed by FACS. Results The lentivirus vector infected the U251 steadily and efficiently. Compared with the control groups, the RNF138-siRNA significantly decreased the expression of RNF138 mRNA for 60%. The U251cell proliferation was inhibited and cell apoptosis increased and the cells were blocked in G2/M phase by knockdown the RNF138 gene. Conclusions The lentivirus vector expressing RNF138-siRNA can significantly decrease the expression of RNF138 ,inhibit the cell proliferation, promote the cell apoptosis, effect the cell cycle and block the cell devision.