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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和治疗作用。方法 本文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、免疫组化、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检测五株亲本胶质瘤细胞表达P2 1WAF1 CIP1 cDNA ,而流式细胞仪未检测到P2 1表达 ;Ad P2 1转导的胶质瘤细胞株均过量表达P2 1;在体内外Ad P2 1能诱导胶质细胞瘤向终末分化、凋亡或诱导炎症反应抑制肿瘤生长或消退。结论 基因转移P2 1WAF1 CIP1 为研究基因治疗恶性脑胶质瘤提供了有效的手段。
Objective To investigate the effect of gene transfer P2 1 WAF1 CIP1 overexpression on human glioma cell apoptosis and its therapeutic effect. Methods We constructed defective recombinant adenoviral vectors: human P2 1 cDNA (Ad P2 1) under the control of the CMV promoter, which encodes the human P2 1 protein. P2 1 expression was detected by flow cytometry and RT-PCR. TUNEL assay, immunohistochemistry, fluorescence microscopy and flow cytometry were used to detect and analyze the apoptosis. Results The mRNA expression of P2 1 WAF1 CIP1 was detected by RT PCR in five parental glioma cells, while P2 1 was not detected by flow cytometry. P2 1 was overexpressed in Ad P2 1 transduced glioma cells. P2 1 can induce glioblastoma to terminal differentiation, apoptosis or induce inflammatory response inhibit tumor growth or regression. Conclusion Gene transfer P2 1 WAF1 CIP1 provides an effective method for gene therapy of malignant gliomas.