摘要
目的观察野生型p16基因对膀胱肿瘤细胞恶性表型及H-ras基因表达的影响。方法构建p16基因逆转录病毒表达载体,转染膀胱肿瘤细胞系EJ;应用Northern杂交检测外源p16基因及H-ras基因表达;流式细胞仪检测细胞周期;双层软琼脂培养检测细胞在体外形成克隆的能力。结果转染p16基因后,肿瘤细胞生长速率降低,体外形成克隆的能力下降,细胞被抑制在G_0+G_1期,细胞内H-rasmRNA表达低于非转染细胞。结论增加细胞内野生型p16基因表达量能抑制突变的H-ras基因表达,从而抑制膀胱癌细胞的恶性增殖。
Objective To investigate the effect of p16 gene transfection on the malignant phenotype and H-ras gene expression of bladder cancer cell line EJ. Methods p16 recombinant retrovirus vector was constructed and transfected into EJ. Northern blot analysis was used to detect the expression of p16 and Hras gene, flow cytometry analysis and soft agar cell culture to detect tumor cell cycle and colony forming ability before and after gene transfection. Results Introduction of p16 gene in EJ significantly suppressed the cell growth and the ability to form colonies in soft agar in vitro. The majority of infected cells were inhibited in G_0+G_1 phases of the cell cycle. H-ras gene expression in transfected cells was significantly reduced as compared with nontransfected cells. Conclusion The increase of wild type p16 gene expression suppressed the malignant proliferation of bladder cancer cells by down regulating the expression of mutant H-ras gene.
出处
《中华实验外科杂志》
CSCD
北大核心
1998年第5期459-460,共2页
Chinese Journal of Experimental Surgery
基金
天津市卫生局科研基金
