siRNA靶向抑制ATDC蛋白对胃癌细胞生物学行为的影响

目的通过比较ataxia—telangiectasiagroupDcomplementinggene（ATDC）在永生化胃黏膜细胞GES与胃癌细胞系MNK45、AGS中的表达差异，探讨其对胃癌细胞生长、增殖及侵袭能力的影响。方法应用Westernblot实验检测ATDC在胃癌细胞系MKN45、AGS及永生化胃黏膜细胞系GES表达差异，利用慢病毒携带的siRNA下调ATDC在MKN45细胞表达水平，采用MTT实验检测细胞生长能力、流式细胞术检测细胞周期、平板克隆实验检测细胞克隆形成能力以及transwell实验检测细胞侵袭能力。结果ATDC在人胃癌细胞系MKN45与AGS细胞中的表达水平均明显高于永生化胃黏膜细胞GES（均P〈0．05）；转染siRNA慢病毒后，与Con—MKN45及MKN45比较，Si—MKN45细胞的表达水平明显下降（P〈0．05）：下调ATDC表达后MKN45细胞的生长能力明显受到抑制（P〈0．05），S期细胞明显减少、增殖指数克隆形成率及侵袭能力均明显降低（均P〈005）。结论慢病毒携带的siRNA下调ATDC表达后能够抑制胃癌细胞MKN45的生长、增殖与侵袭能力，可为胃癌的基因治疗提供新的靶点。

Effects of ATDC gene silencing by siRNA on proliferation and migration of gastric cancer cells CA/ Weilong,WANG Weimin,PAN Jie,et al. Department of General Surgery,Huzhou Central Hospital,Huzhou 313000, China

Objective TO investigate the effect of ATDC gene silencing by siRNA on the proliferation and migration of gastric cancer cells. Methods The expression of ataxia-telangiectasia group D complemenfing（ATDC） was detected in immor- talized gastric mucosal epithelium cell line GES and gastric cancer cell lines MKN45, AGS by Western blot. Lentivirus-mediated siRNA targeting ATDC was transfected into MKN45 cells, and western blot was performed to evaluate the inhibitory effect of siR- NA on ATDC expression. The proliferation, cell cycle, growth potentiality and migration of MKN45 cells were evaluated by MTT, plate cloning formation test, flow cytometry and transwell assays, respectively. Results The expression of ATDC was signifi- cantly higher in both MKN45 and AGS cells compared with GES cells. Lentivirus-mediated siRNA targeting ATDC markedly re- pressed the expression of ATDC in MKN45 cells. MTT and plate cloning formation assays showed that down-regulation of ATDC significantly inhibited the proliferation and growth of MKN45 cells （P〈0.05）. Flow cytometry showed that the number of cells transfected with ATDG siRNA had higher G1, lower S phase and lower proliferation index （PI）（P〈0.05） than those of controls. Transwell assays revealed that the migration of cells transfected with ATDC siRNA was significantly deceased compared with controls. Conclusion The expression of ATDC is increased in gastric cancer cells, and silencing of ATDC significantly sup- presses the proliferation and migration of gastric cancer cells, indicating that ATDC may be a novel target for gene therapy of gastric cancer.