摘要
目的 探讨复制缺陷型腺病毒介导的 p16基因转染对人膀胱癌细胞生长的影响。 方法 将 p16重组腺病毒 (Ad p16 )感染p16蛋白表达阴性的人膀胱移行细胞癌T2 4细胞株 ,Ad LacZ、X gal染色法检测重组腺病毒的转染效率 ;Westernblot法检测 p16蛋白的表达 ;MTT法及流式细胞术评估Ad p16对T2 4细胞增殖的影响。 结果 在感染复数 (MOI)为 5 0时 ,重组腺病毒对T2 4细胞的转染率达 97% ;此Ad p16在T2 4细胞中可获高效表达 ;与转染Ad LacZ组及未转染组相比 ,转染Ad p16组T2 4细胞生长受到明显抑制 (P <0 .0 5 ) ,细胞周期分析表明生长抑制主要发生在G1 S节点。 结论 腺病毒载体可有效地将外源 p16基因导入T2 4细胞 ;p16基因重组腺病毒载体转染能抑制T2 4细胞的生长。
Objective To investigate the effects of full length p16 gene transfer by replication-defective recombinant adenoviruses on cell growth of the human bladder carcinoma. Methods Ad-p16 was infected into the human bladder carcinoma cell line with no expression of p16 protein,T24.The transfection efficiency of the recombinant adenoviruses was tested using Ad-LacZ by X-gal staining.The expression of p16 protein was evaluated by Western-blot analysis. The effects of Ad-p16 infection on cell proliferation were evaluated by MTT assay and flow cytometry. Results Higher transfection efficiency(97%) to T24 could be obtained at MOI(multiplicity of infection) of 50.This Ad-p16 has a high gene expression in T24 cells.The proliferation of Ad-p16 infected T24 cells was significantly suppressed as compared with that of Ad-LacZ-infected and non-infected cells ( P <0.05).The inhibition of cell growth occurred mainly at G1-S checkpoint through flow cytometry analysis. Conclusions Foreign p16 gene could be transfected effectively into cultured T24 with the recombinant adenoviruses.The proliferation of T24 cells could be inhibited by adenovirus-mediated p16 gene therapy which might play a potential role in human bladder carcinoma treatment.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2002年第7期434-436,共3页
Chinese Journal of Urology
