摘要
目的 研究降低c Ha ras癌基因表达对膀胱癌细胞生长和粘附的影响。方法 将反义c Ha ras基因导入膀胱癌细胞系TBC 1细胞株 ,筛选阳性细胞克隆。Southern杂交鉴定外源基因整合情况 ,荧光分光光度法检测c Ha ras表达 ,绘制生长曲线 ,体外检测细胞粘附率。结果 筛选出细胞克隆TBC neo和TBC antiras ,放射自显影显示只有TBC antiras有外源性c Ha ras杂交带 ,亲本细胞TBC 1和对照细胞TBC neo只有内源性杂交带 ,与TBC 1(39.7± 6 .2 ,32 .8% )和TBC neo(43.2±5 .7,35 .7% )相比TBC antiras细胞的c Ha ras蛋白水平 (2 5 .6± 4.2 )及粘附率 (16 .4% )明显降低 (P<0 .0 5 )。生长曲线显示TBC antiras生长受到抑制。结论 降低c Ha ras癌基因表达抑制膀胱癌细胞生长和粘附 ,针对ras癌基因或癌蛋白的药物可为膀胱癌的临床治疗提供新途径。
Objective To investigate the effects of decreasing expression of c Ha ras on the growth and adhesion of human bladder papillary tumor. Methods Antisense c Ha ras gene was transferred into bladder papillary tumor cell line (TBC 1), then positive cells were selected. Exogenous gene integration was identified by using Southern Blot,c Ha ras gene expression detected by flurospectrophotometry, growth curve drawn and tumor cell adhesion in vitro measured. Results The cells transfected pSV2neo or anti raspSV2neo vectors had been cloned. Only TBC antiras had exogenous c Ha ras blot, whereas native cell TBC 1 and control cell TBC neo had endogenous ones on autoradiographs. Expression of c Ha ras protein and adhesion ratio in TBC antiras cell were significantly decreased as compared with those of TBC 1 and TBC neo(25.6±4.2 vs 39.7±6.2 and 42.3±5.7,13.4% vs 32.8% and 35.7%,P<0.05). The growth curve showed that the growth of TBC antiras was inhibited.Conclusion Decreasing expression of c Ha ras gene inhibited TBC 1 growth and adheion.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2000年第2期178-179,共2页
Chinese Journal of Experimental Surgery
