重组腺病毒介导P21^(WAF1/CIP1)诱导人脑胶质瘤细胞凋亡的体外实验研究

Experimental study of the gene transfer P21 WAF1/CIP1 induced apoptosis of glioma cells in vitro

目的 :研究基因转移P2 1WAF1/CIP1过量表达对人脑胶质瘤细胞凋亡的影响。方法 :构建缺陷型重组腺病毒载体 :在CMV启动子控制下的人p2 1cDNA(Ad P2 1) ,其编码人P2 1蛋白。通过流式细胞仪、RT PCR进行P2 1表达检测 ;应用TUNEL法、荧光显微镜和流式细胞仪进行相关凋亡检测、分析。结果 :RT PCR检测 5株亲本胶质瘤细胞表达p2 1cDNA ,而流式细胞仪未检测到P2 1表达 ;Ad P2 1转染的胶质瘤细胞株均过量表达P2 1;在体外通过细胞形态学、分子生物学等检测均证实Ad P2 1能诱导胶质瘤细胞向终末分化并诱导细胞凋亡。结论 :基因转移P2

Objective To examine the apoptotic effects of overexpression of P21 WAF1/CIP1 on human malignant glioma cells in vitro.Methods A recombinant advenoviral vector(AdP21) containing human p21 cDNA was constructed under the control of cytomegalovirus(CMV) promotor,which encoded human P21 cyclin dependent kinase inhibitor.P21 WAF1/CIP1 expression was assessed by flow cytometric analysis and RTPCR,and the apoptosis of glioma cells was detected by TUNEL assay and flow cytometric analysis.Results p21 cDNA was detected in five glioma cells by RTPCR,however it was undetectable by flow cytometric analysis,while tumor cells transducted by AdP21 over expressed P21.Analytic methods of cell morphology and molecular biology in vitro confirmed that AdP21 can induce glioma cells growth arrest and apoptosis.Conclusion The effect of gene transfer P21 inducing the apoptosis suggests it can be widely used as a tumoricidal agent instead of a tumoristatic agent in preventing gliomas.