Survivin基因沉默抑制胃癌MGC-803细胞增殖并增强其对塞来昔布的.DOC

Survivin基因沉默抑制胃癌MGC-803细胞增殖并增强其对塞来昔布的敏感性 方雷1 吴海滨2 陈晓岗1 （宁波第一医院宁波宁波urvivin基因沉默对人胃癌MGC-803细胞增殖和对化疗药物塞来昔布敏感性的影响。方法 设计合成survivin的siRNA序列，Lipofectamine?2000 转染入MGC-803细胞。采用RT-PCR和Western blot检测survivin在干扰后mRNA和蛋白的表达情况，利用流式细胞仪检测细胞周期。通过MTT法和细胞克隆形成试验法察survivinurvivin基因沉默48h后，MGC-803细胞的survivin基因和蛋白表达明显降低，差异有统计学意义(P0.05)。细胞周期被阻滞在G0/G1期，S期细胞数减少；差异有统计学意义(P0.05)。survivin基因沉默组细胞对塞来昔布的敏感性显著性增强(P0.05)。结论Survivin特异性siRNA能显著沉默MGC-803细胞survivin基因，抑制细胞增殖，并增强MGC-803细胞对塞来昔布的敏感性。 关键词 ：Survivin； siRNA； 胃癌；MGC-803； 细胞增殖 ；塞来昔布 Survivin gene silencing Inhibited Proliferation of Human gastric cancer cell line MGC-803 and Enhanced Their Sensitivity to Celecoxib. Fang Lei 1 Wu Hai-bin2 Chen Xiao-Gang 1 (Ningbo First Hospital1, General Surgery, 315 010, Ningbo, P.R. China)(Ningbo University School of Medicine 2, 315010, Ningbo, P.R. China) 【Abstract】 Objective This study investigated the effect of small interfering RNA-mediated urvivin knock-down on proliferation of human gastric cancer cell line MGC-803 and their sensitivity to Celecoxib. Methods The siRNA against Survivin was constructed and transfected into MGC-803 cells with Lipofectamine? 2000. The expression of Survivin was detected by RT-PCR and Western blot. Flow cytometry was used to detect the cell cycle. Sensitivity to Celecoxib after transfection were examined by MTT assay and clonogenic assay. Results In MGC-803 cells, the protein and mRNA levels of Survivin were decreased significantly after transfection, and reduction of proliferation was related to an increase in the fraction of Go/G1 phase. The sensitivity of MGC-803 cells to Celecoxib was increased significantly after transfection. Conclusion The Survivin specia1 siRNA silenced Survivin，decreased MGC-803 cells proliferation and enhanced their sensitivity to Celecoxib. Key words: Survivin；siRNA；gastric cancer; MGC-803；Cell proliferation; Celecoxib 胃癌是最常见的消化系统肿瘤之一，对人们健康照成很大危害[1]。本文通过将合成的siRNA转染入胃癌MGC-803细胞，分析siRNA对其Survivin基因沉默的效率；Survivin基因被沉默后，肿瘤细胞周期的变化及其对塞来昔布的敏感性的影