EGFL6基因沉默对人黑素瘤细胞增殖及侵袭的影响

Effects of EGFL 6 gene silencing on proliferation and invasion of human melanoma cells

目的 ：通过特异性小干扰RNA（small interference RNA,si RNA）抑制人黑素瘤A375细胞中表皮生长因子样结构域蛋白6（epidermal growth factorlike-domain 6,EGFL 6）基因表达,探讨EGFL 6基因沉默对人黑素瘤细胞增殖及侵袭的影响。方法：设计合成靶向EGFL 6基因的siRNA（命名为EGFL6-siRNA）,通过脂质体转染法将该siRNA转染入黑素瘤A375细胞,然后采用实时荧光定量PCR法和蛋白质印迹法分别检测黑素瘤A375细胞中EGFL6 mRNA及蛋白的表达水平,细胞计数试剂盒-8（cell counting kit-8,CCK-8）法和Transwell实验分别检测A375细胞增殖和侵袭能力的变化。结果：在成功转染EGFL6-siRNA 48 h后,A375细胞中EGFL6 mRNA及蛋白表达水平均明显下降（P值均〈0.01）。EGFL6-siRNA转染组A375细胞在转染后24~72 h各时间点的细胞增殖能力均受到明显抑制（P值均〈0.01）,且细胞侵袭能力显著降低（P〈0.01）。结论：特异性siRNA能够有效地沉默人黑素瘤A375细胞中EGFL 6基因的表达,并抑制肿瘤细胞的增殖和侵袭。

Objective： To explore the effects of epidermal growth factor-like domain 6（EGFL 6） gene silencing by specific small interference RNA（siRNA） on the proliferation and invasion of human melanoma cells.Methods： The double-strand siRNA targeting EGFL 6 gene（named as EGFL6-siRNA） was designed and synthesized. The EGFL6-siRNA was transfected into human melanoma A375 cells by liposome transfection method. The mRNA and protein expression levels of EGFL6 in A375 cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The proliferation and invasion abilities of A375 cells were analyzed by cell counting kit-8（CCK-8） and Transwell assay, respectively.Results： After EGFL6-siRNA was successfully transfected into human melanomaA 375 cells for 48 h, the EGFL6 mRNA and protein expression levelsin A375 cells were significantly decreased（both P 〈 0.01）. The EGFL6-siRNA could signif cantly inhibit the proliferation of A375 cells at different time points（24-72 h） after transfection（all P 〈 0.01）, and signif cantly inhibit the invasion of A375 cells（P 〈 0.01）.Conclusion： The specific siRNA can effectively silence EGFL 6 gene expression in human melanoma A375 cells, and suppress the proliferation and invasion of melanoma cells.