重组腺病毒介导P21WAF1/CIP1诱导人脑胶质瘤细胞凋亡的体外实验研究-东南大学学报医学版

重组腺病毒介导P21WAF1/CIP1诱导人脑胶质瘤细胞凋亡的体外实验研究

单位：1.东南大学附属中大医院,神经外科,江苏,南京,210009； 2.苏州大学第一附属医院,神经外科,江苏,苏州,215006

分类号：Q255

出版年·卷·期（页码）：2002·21·第四期（289-292）

摘要：

目的:研究基因转移P21WAF1/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p21 cDNA(Ad-P21),其编码人P21蛋白.通过流式细胞仪、RT-PCR进行P21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p21cDNA,而流式细胞仪未检测到P21表达;Ad-P21转染的胶质瘤细胞株均过量表达P21;在体外通过细胞形态学、分子生物学等检测均证实Ad-P21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P21WAF1/CIP1为研究基因治疗恶性脑胶质瘤提供了有效的手段.

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.

参考文献：

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