RNA干扰技术干涉后核糖体蛋白S15a在胃癌中的表达及意义

摘要

Objective To investigate the effects of ribosomal protein S15a (RPS15a) in proliferation of gastric cancer cellBGC823 by RNA interference technique. Methods BGC823 cells were transfected with random non- coding sequences (siNC group), siRPS15a- 1 (siRPS15a- 1 group) or siRPS15a- 2 (siRPS15a- 2 group), respectively. RPS15a RNA and protein expression was detected by RT- PCR and Western Blot test, respectively;and the cellproliferation was determined by flow cytometry. Re-sults The expression levels of RPS15 in SiRPS15a- 1 and siRPS15a- 2 groups were significantly lower than those in non- trans-fected BGC823 cells (WT group) and siNC group (al P<0.05); no significant differences was found between WT and siNC groups (P>0.05). cellproliferation rate were decreased significantly in SiRPS15a- 1 and siRPS15a- 2 groups, and most cells were in G0/G1 phase. Conclusion RPS15a RNA interference technique inhibits cells growth, which is associated with the knock- down of RPS15a expression in BGC823 cells.%目的：探讨RNA干扰（RNAi）技术干涉后核糖体蛋白S15a（RPS15a）在胃癌中的表达及意义。方法选取12例胃癌患者手术切除的胃癌组织，利用RNAi干涉RPS15a基因，将小片段克隆到质粒中，分别提取质粒后转染胃癌细胞BGC823。将未转染、随机小片段转染非编码序列、siRPS15a-1转染、siRPS15a-2转染的BGC823细胞分为BGC823- WT组、siNC组、siRPS15a-1组、siRPS15a-2组，RT- PCR法及Western blot分别检测各组RPS15a RNA及蛋白的表达情况，流式细胞仪检测细胞增殖的变化。结果 RNAi后细胞中出现了明显的绿色荧光，提示此转染方式是可行的。转染检测结果表明，本实验提取的RNA未受到质粒DNA的污染，可以进行表达的比较。siRPS15a-1组、siRPS15a-2组RNA及蛋白表达相对灰度比值与BGC- WT组、siNC组比较，差异均有统计学意义（均P＜0.05），BGC- WT组与siNC组比较差异无统计学意义（P＞0.05）。RNAi后细胞增殖速度显著下降，多数细胞集中在G0/G1期。结论 RNAi技术不仅可以下调RPS15a在胃癌中表达，而且还可以抑制胃癌细胞的增殖活性。