siRNA沉默脂肪酸合成酶基因表达对食管癌细胞增殖的影响

Effect of silencing fatty acid synthase gene(FASN)by siRNA interference on proliferation of human esophageal cancer cells

目的观察小干扰RNA(siRNA)沉默脂肪酸合成酶基因(FASN)表达对食管癌细胞增殖的影响。方法将化学合成的FASN的小干扰siRNA用脂质体Lipofectamine2000转染人食管癌细胞株TE13细胞,采用的反转录-聚合酶链反应(RTPCR)检测FASN mRNA的表达、Western blot方法检测siRNA干扰后FASN蛋白的表达及与细胞增殖相关的蛋白cyclin D1(CCND1)的表达情况、CCK-8检测细胞增殖、Transwell实验检测细胞迁移能力。结果转染了FASN-siRNA的食管癌TE13细胞,RT-PCR及Western blot结果显示FASN在基因及蛋白水平明显下调(P<0.05);与对照组比较,细胞增殖受到抑制,与增殖相关的蛋白CCND1表达下调(P<0.05);FASN干扰后明显抑制了人食管癌细胞株TE13细胞迁移能力(P<0.05)。结论应用FASN-siRNA体外抑制食管癌细胞FASN的表达,从而抑制食管癌细胞增殖,抑制了细胞迁移,靶向FASN可以作为食管癌治疗的一个新的靶点。

Objective To investigate the effect of silencing fatty acid synthase gene （FASN） by siRNA interference on proliferation of human esophageal cancer cells. Methods Chemically syn- thesized siRNA targeting FASN（FASN-siRNA） was transfected into transfected into human esophageal cancer cell line TEl3 ceils by lipofectamine2000. FASN mRNA was measured by reverse transcription Polymerase chain reaction（ RT-PCR）, and the expression of FASN, cyclin D1 （CCND1） were detec- ted by Western blotting. Proliferation of TEl3 ceils was determined by CCK-8 assay. The activities of motility of TEl3 cells were assessed by celhranswell chamber invasion assay in vitro. Results In the FASN-siRNA group, the FASN mRNA and protein levels were down-regulated remarkably （ P 〈 0.05 ）. The abilities of proliferation were inhibited and level of CCND1 was down-regulated （P 〈 0.05 ）. The motility of TEl3 cells were inhibited significantly in vitro （P 〈 0.05 ）. Conclusion Silencing FASN could regulate proliferation of human esophageal cancer cells. Targeting FASN may be a new therapeu- tic strategy for human esophageal carcinoma.