siRNA干扰FLOT2基因表达对肾细胞癌细胞增殖的影响

Effect of silence of FLOT2 expression by siRNA on proliferation of human renal cell carcinoma cells

目的:探讨FLOT2在肾细胞癌细胞株中的表达,并观察小干扰RNA沉默FLOT2基因对肾细胞癌786-O细胞增殖能力的影响。方法:将化学合成的FLOT2的小干扰siRNA用脂质体Lipofectamine2000转染至肾细胞癌786-O细胞;分别用RT-PCR、Western blot方法检测细胞转染前后FLOT2以及细胞增殖相关因子CCND1的表达。CCK-8与克隆形成实验检测其增殖情况。结果:转染FLOT2-siRNA的肾细胞癌786-O细胞,RT-PCR、Western blot检测显示FLOT2在基因及蛋白水平明显下调(P<0.05);与对照组比较786-O细胞增殖能力明显下降(P<0.05),细胞增殖相关因子CCND1明显下调(P<0.05)。结论:FLOT2在肾细胞癌细胞系中高表达,采用特异性FLOT2-siRNA能够显著降低FLOT2和CCND1表达。干扰FLOT2表达能显著抑制786-O细胞的增殖,FLOT2基因可能参与了肾细胞癌的生物学行为。

Objective： To investigate the effects of flotillin-2（ FLOT2） gene expression silencing on the proliferation in vitro inhuman renal cell carcinoma cell line 786-O. Methods： Chemically synthesized siRNA targeting FLOT2（ FLOT2-siRNA） was transfected into human renal cell carcinoma cell line 786-O cells by lipofectamine 2 000.The expressions of FLOT2 mRNA were detected by PCR. The proteins of FLOT2 and CCND1 and after cell transfection were detected by Western blot. The effects of altered expression of FLOT2 on the cell proliferation were measured by CCK-8 and colony formation assay. Results： After cell transfection,the FLOT2 mRNA expressions and proteins were significantly decreased（ P〈0. 05）. The abilities of proliferation were inhibited and level of CCND1 was down-regulated（ P〈0. 05）. Conclusion： FLOT2 expression in renal clear cell carcinoma is higher than in normal renal cells,FLOT2-siRNA can down-regulate the expression of FLOT2 protein in 786-O cells,inhibit the proliferation and colony formation of 786-O cells effectively,which suggest that FLOT2 may be involved in the malignant behavior of renal cell carcinoma.