RB_1抑癌基因对人膀胱癌细胞抑癌作用的研究

A study of tumor suppression effect of RB_1 gene on human bladder carcinoma cells in vitro

采用电穿孔方法将组装于ＤＮＡ质粒载体ｐＣＭＶｎｅｏ的ＲＢ１基因导入该基因功能丧失的膀胱癌细胞系ＨＴＢ９－５６３７。结果发现，被ＲＢ１基因转染的膀胱癌细胞的生长速率减低，半固体培养集落形成能力消失，接种裸鼠无致瘤性，流式细胞术（ＦＣＭ）检测细胞分裂各相所占比例分布明显改变，Ｎｏｒｔｈｅｒｎ印迹分析发现其Ｈａ－ｒａｓ癌基因表达受到一定抑制。结果表明ＲＢ１抑癌基因对人类膀胱癌细胞具有明显抑癌作用。

By electroporation,the reconstructed RB1 expression DNA plasmid vector,pCMV-neo-RB1,was introduced into a bladder carcinoma cell line,HTB9-5637.The experiments performed on the transfected cells showed that the growth rate of these cells was significantly retarded and the abilities of anchorage independent growth and tumorigenicity in nude mice were lost.Flowcytometry measurement indicated that the proportion of cyclical S-phase cells among the transftetants was distinctly lessened. Northern blot analysis of Ha-ras oncogene revelaled that its expression in transfected cells was markedly reduced.These results whow that the RB1 gene can work as a tumor suppressor gene in human bladder carcinoma cells as in other cancer cells.