PinX1基因过表达对神经胶质瘤细胞增殖的影响  

Efef cts of PinX1 overexpression on the pr oliferation of glioma cells

作　　者：杜颖[1] 李中林[1] 程乾[1] 曹梦函[1] 梅鹏金[1] 白津[1] 郑骏年[1] 

机构地区：[1]徐州医学院江苏省肿瘤生物治疗重点实验室,江苏徐州221002

出　　处：《徐州医学院学报》2015年第10期639-643,共5页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金（81472663,81201636）;江苏省自然科学基金（BK2012139）;江苏省高校自然科学基金（13KJB320028）;徐州市科技局项目（XM138075）

摘　　要：目的：探讨PinX1基因过表达对神经胶质瘤细胞株增殖的影响及其分子作用机制。方法利用pEG-FP－C3－PinX1过表达质粒和pEGFP－C3对照组质粒，分别转染神经胶质瘤U87和U251细胞，CCK8细胞增殖实验分析PinX1过表达后对2种神经胶质瘤细胞增殖的影响，流式细胞仪分析2种神经胶质瘤细胞周期的变化， Western blot检测PinX1蛋白c、yclin 家族蛋白、CDK抑制剂蛋白及人端粒酶逆转录酶（ hTERT）的表达水平。结果转染pEGFP－C3－PinX1过表达质粒可以有效增加神经胶质瘤U87和U251细胞中PinX1蛋白的表达，PinX1过表达后抑制细胞的增殖，PinX1过表达后下调hTERT的表达，提高 p27的表达，降低cyclin D1和cyclin E的表达，使细胞周期停滞在G1期。结论 pEGFP－C3－PinX1过表达质粒可以有效地提高神经胶质瘤U87和U251细胞中PinX1的表达，PinX1过表达后可能通过影响cyclins、CDK抑制剂蛋白以及hTERT的表达从而使细胞周期停滞在G1期，最终显著抑制神经胶质瘤细胞的增殖。Objecit ve To investigate the effects of PinX1 overexpression on the proliferation of U87 and U251 glio-ma cells and related molecular mechanisms.Metho ds The plasmid pEGFP-C3-PinX1 or pEGFP-C3 was transfect-ed into U87 and U251 glioma cells using Lipofectamine 2000 reagent.Then, the effects of up-regulated PinX1 on the proliferation of U87 and U251 glioma cells were detected by CCK-8 assay.Meanwhile, changes in cell cycle were meas-ured by flow cytometry.The levels of PinX1, cyclins, CDK inhibitor proteins and human telomerase reverse transcriptase （hTERT） were determined by Western blot.Resul ts The plasmid pEGFP-C3-PinX1 could remarkably enhance the qualities of PinX1 in both U87 and U251 glioma cell lines.Up-regulation of PinX1 dramatically blocked the proliferation of the cells, while reducing the amounts of cyclin D1, cyclin E and hTERT, in spite of an increase in p27 expression. The cell cycle arrested in the G1phase .Conclusions Overexpression of PinX1 can effectively enhance the levels of cyc-lins, CDK inhibitor proteins and hTERT, so as to arrest glioma cells in the G1 phase and finally affect their proliferation.

关 键 词：PinX1 神经胶质瘤 增殖 人端粒酶逆转录酶 

分 类 号：R73-362[医药卫生—肿瘤]