人Ras同源物基因家族成员B真核表达载体的构建及对肾癌细胞786—0增殖和凋亡的影响

Construction of eukaryotic expression vecter for Ras homolog gene family members B and its effecton proliferation and apoptosis of renal cell carcinoma cell line 786-0

目的观察人Ras同源物基因家族成员B（RhoB）对肾癌细胞株786一O细胞增殖和凋亡的影响。方法构建pcDNA3．0-Flag—RhoB真核表达载体，脂质体转染法将其转入肾癌细胞786一O，利用Westernblot检测细胞中RhoB的表达。用MTS法和流式细胞仪检测RhoB对肾癌细胞增殖和凋亡效应的影响。结果真核表达载体pcDNA3．0-Flag-RhoB构建成功，脂质体转染后RhoB在786一O细胞中表达明显升高；在转染后48、72、96h，重组质粒组细胞的增殖速度明显低于空载体组和阴性对照组（P〈0．05）；转染重组质粒细胞凋亡率为（24．05±0．60）％，明显高于空载体组[（1．67±0．30）％]和阴性对照组[（1．22±0．50）％，P〈0．05]。结论RhoB对肾癌细胞具有抑制增殖和诱导凋亡的作用。

Objective To study the effect of Ras homolog gene family members B （RhoB） on pro- liferation and apoptosis of renal cell carcinoma cell line 786-0. Methods The eukaryotic expression vector pcDNA3. 0-Flag-RhoB was constructed and transfected into 786-0 cells using liposome-mediate method. RhoB protein level in 786-0 cells was detected by using Western blotting. The effects of RhoB on prolifera- tion and apoptosis of 786-0 ceils were analyzed by using MTS method and flow cytometry. Results The eukaryotic expression vector pcDNA3. O-Flag-RhoB was successfully constructed. Western blotting indica- ted that RhoB was highly expressed in transfected 786-0 cells. At 48, 72 and 96 h after transfection, pro- liferation of 786-0 cells was significantly inhibited in the recombinant plasmid group as compared with the empty vector group and negative control group （ P 〈 0. 05 ）. The apoptosis rate in the recombinant plasmid group was （ 24.05± 0. 60） %, significantly higher than in the empty vector group [ （ 1.67±0. 30） % ] and negative control group [ （ 1.22±0. 50） % ] （P 〈 0.05 ）. Conclusion RhoB can inhibit the proliferation and induce aoootosis of renal cell careinoma cells.