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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 study the effect of reducing oncogene expression of c Ha ras on the growth and adhesion of bladder cancer cells. Methods Antisense c Ha ras gene was transfected into bladder cancer cell line TBC 1 and the positive clones were screened. Southern hybridization was used to identify the integration of exogenous genes. Fluorescence spectrophotometry was used to detect the expression of c Ha ras and the growth curve was drawn. Cell adhesion rate was detected in vitro. Results TBC neo and TBC antiras were screened out by autoradiography. Only TBC antiras had exogenous c Ha ras hybridization, TBC 1 parental and TBC neo only had endogenous hybridization with TBC 1 (39.7 ± 6,22, 32.8%) compared with TBC neo (43.2 ± 5 .7, 35.7%). The level of c Ha ras protein in TBC antiras cells (25.6 ± 4.2) 4%) was significantly lower (P <0. 05). Growth curves showed that TBC antiras growth was inhibited. Conclusions Reducing the expression of c Ha ras oncogene inhibits the growth and adhesion of bladder cancer cells, and the drugs targeting ras oncogene or oncoprotein may provide a new approach for the clinical treatment of bladder cancer.