浙江医学
浙江醫學
절강의학
ZHEJIANG MEDICAL JOURNAL
2014年
15期
1310-1313,1320
,共5页
刘勇攀%石定%王泽军%于香
劉勇攀%石定%王澤軍%于香
류용반%석정%왕택군%우향
核糖体蛋白%S15a%胃癌%RNA干扰
覈糖體蛋白%S15a%胃癌%RNA榦擾
핵당체단백%S15a%위암%RNA간우
Ribosomal protein S15a(RPS15a)%Gastric cancer (GC)%RNA interference (RNAi)
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。
目的:探討RNA榦擾(RNAi)技術榦涉後覈糖體蛋白S15a(RPS15a)在胃癌中的錶達及意義。方法選取12例胃癌患者手術切除的胃癌組織,利用RNAi榦涉RPS15a基因,將小片段剋隆到質粒中,分彆提取質粒後轉染胃癌細胞BGC823。將未轉染、隨機小片段轉染非編碼序列、siRPS15a-1轉染、siRPS15a-2轉染的BGC823細胞分為BGC823- WT組、siNC組、siRPS15a-1組、siRPS15a-2組,RT- PCR法及Western blot分彆檢測各組RPS15a RNA及蛋白的錶達情況,流式細胞儀檢測細胞增殖的變化。結果 RNAi後細胞中齣現瞭明顯的綠色熒光,提示此轉染方式是可行的。轉染檢測結果錶明,本實驗提取的RNA未受到質粒DNA的汙染,可以進行錶達的比較。siRPS15a-1組、siRPS15a-2組RNA及蛋白錶達相對灰度比值與BGC- WT組、siNC組比較,差異均有統計學意義(均P<0.05),BGC- WT組與siNC組比較差異無統計學意義(P>0.05)。RNAi後細胞增殖速度顯著下降,多數細胞集中在G0/G1期。結論 RNAi技術不僅可以下調RPS15a在胃癌中錶達,而且還可以抑製胃癌細胞的增殖活性。
목적:탐토RNA간우(RNAi)기술간섭후핵당체단백S15a(RPS15a)재위암중적표체급의의。방법선취12례위암환자수술절제적위암조직,이용RNAi간섭RPS15a기인,장소편단극륭도질립중,분별제취질립후전염위암세포BGC823。장미전염、수궤소편단전염비편마서렬、siRPS15a-1전염、siRPS15a-2전염적BGC823세포분위BGC823- WT조、siNC조、siRPS15a-1조、siRPS15a-2조,RT- PCR법급Western blot분별검측각조RPS15a RNA급단백적표체정황,류식세포의검측세포증식적변화。결과 RNAi후세포중출현료명현적록색형광,제시차전염방식시가행적。전염검측결과표명,본실험제취적RNA미수도질립DNA적오염,가이진행표체적비교。siRPS15a-1조、siRPS15a-2조RNA급단백표체상대회도비치여BGC- WT조、siNC조비교,차이균유통계학의의(균P<0.05),BGC- WT조여siNC조비교차이무통계학의의(P>0.05)。RNAi후세포증식속도현저하강,다수세포집중재G0/G1기。결론 RNAi기술불부가이하조RPS15a재위암중표체,이차환가이억제위암세포적증식활성。
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.