节律基因Bmal1对胃癌细胞BGC-823增殖及凋亡的影响

Effects of Bmal1 on proliferation and apoptosis of gastric cancer cells BGC-823

目的探讨节律基因Bmal1对胃癌细胞BGC-823增殖及凋亡的影响。方法构建针对Bmal1序列的shRNA,用脂质体介导转染人胃癌细胞BGC-823,G418筛选稳定细胞株(转染组),同时设对照组(空白组)。用克隆形成率检测细胞生长,用流式细胞术检测细胞凋亡及细胞周期,RT-PCR及Western blot法检测相关基因及蛋白表达。结果转染组克隆形成率低于对照组(P<0.05)。与对照组比较,转染组Bmal1总凋亡率升高(P<0.05),其细胞周期再分布出现G1期缩短(P<0.05),G2期延长(P<0.05),同时发现Bcl-2 mRNA表达下调(P<0.05),c-Myc mRNA表达上调(P<0.05)。结论 Bmal1可以抑制胃癌细胞BGC-823增殖,促进细胞凋亡,有可能成为诱导胃癌细胞凋亡的新靶点。

Objective To investigate the effects of Bmall on the proliferation and apoptosis of gastric cancer cells BGC-823. Methods The shRNA targeting Bmall sequencing was constructed. The recombination pGCsilencerTM-Bmall （-） and pGCsilencerTM-Bmall （ ＋ ） plasmids were purchased and transfected into BGC-823 by lipofectamine 2000. The stable transfected cells were selected by G418 as the transfection group, the control group without any treatment. The cell growth activity was determined by colony formation assay. The apoptosis and cell cycle were detected by flow cytometry. The related genes and protein expressions were evaluated by RT-PCR and Western blot. Results The cloning efficiency in the transfection group was lower than that in the control group （P 〈 0.05 ）. Compared with the control group, the total ap- optosis rates of Bmall in the transfection group were increased （P 〈 0.05 ）, and the cell cycle redistribution indicated that suppressing Bmall expression in BGC823 decreased G1 phase （41.58%） and increased G2 phase （31.8% ）. meanwhile, the expression level of Bcl-2 mRAN was down-regulated （ all P 〈 0.05 ）, while the expression level of c-Myc mRNA was up-regulated significantly （P 〈 0.05 ）. Conclusion Bmall can inhibit the proliferation of gastric cancer cells, promote apoptosis, and it might be a new target for inducing apoptosis of gastric cancer cells.