ABCE1基因沉默抑制T24细胞增殖并增强其对吉非替尼的敏感性.docVIP

ABCE1基因沉默抑制T24细胞增殖并增强其对 吉非替尼的敏感性 滕立新 沈阳医学院奉天医院泌尿外科 110024 摘要：目的 探讨沉默ABCE1基因对人膀胱癌T24细胞增殖和对吉非替尼化疗敏感性的影响。方法 设计合成ABCE1的siRNA序列，LipofectamineTM2000 转染入T24细胞。采用RT-PCR和Western blot法检测基因沉默后ABCE1mRNA和蛋白的表达情况，采用流式细胞仪技术检测T24细胞周期。通过MTT检测和细胞克隆形成试验察测沉默ABCE1基因后T24细胞对吉非替尼的敏感性。结果 沉默ABCE1基因48h后，T24细胞ABCE1mRNA及蛋白表达明显降低，差异有统计学意义(P0.05)。T24的细胞周期被阻滞于G0/G1期，且S期的细胞数减少；差异有统计学意义(P0.05)。沉默ABCE1基因后T24细胞对吉非替尼的敏感性显著性增强。结论ABCE1特异性siRNA能显著沉默T24细胞ABCE1基因表达，抑制T24细胞增殖，并增强其对吉非替尼的敏感性。 关键词 ：ABCE1；siRNA；膀胱癌；T24细胞；细胞增殖 ；吉非替尼 ABCE1 gene silencing Inhibited Proliferation of Human bladder cancer Cell T24 and Enhanced Their Sensitivity to Geftinib Teng Li-xin Shenyang Medical College Fengtian Hospital Department of Urology Shenyang 110024 【Abstract】 Objective To investigate the effect of small interfering RNA-mediated ABCE1 knock-down on proliferation of human bladder cancer Cell line T24 and their sensitivity to Geftinib. Methods The siRNA against ABCE1 was constructed and transfected into T24 cells with Lipofectamine? 2000. The expression of ABCE1 was detected by RT-PCR and Western blot. Flow cytometry was used to detect the cell cycle. Sensitivity to Geftinib after transfection were examined by MTT and clonogenic assay. Results In T24 cells, the protein and mRNA levels of ABCE1 were decreased significantly after transfection, and reduction of proliferation was related to an increase in the fraction of Go/G1 phase. The sensitivity of T24 cells to Geftinib was increased significantly after transfection. Conclusion The ABCE1 specia1 siRNA silenced ABCE1，decreased T24 cells proliferation and enhanced their sensitivity to Geftinib. Keywords: ABCE1; siRNA; bladder cancer; T24 cells; cell proliferation; Geftinib 膀胱癌是临床泌尿系最常见的恶性肿瘤，我国占泌尿系统恶性肿瘤发病率的也是全身十大常见肿瘤之一。可发生于膀胱的各层组织ABCE1基因的特异性siRNA，将其转染至人膀胱癌T24细胞沉默ABCE1基因， 探讨siRNA转染T24对ABCE1基因沉默的效率及对细胞凋亡和细胞周期的影响，以及ABCE1基因沉默后对T24细胞对吉非替尼(Geftinib)敏感性的影响。为探讨RNA干扰(RNAi)技术抑制ABCE1活性对人膀胱癌基因的影响以及做为增敏药物提供理论依据。 1　材料和方法 1.1主要材料 膀胱癌T24细胞株