EGFL6基因沉默对人黑素瘤细胞增殖及侵袭的影响

发布时间：2018-04-27 16:48

本文选题：黑素瘤 + RNA干扰　；参考：《肿瘤》2015年04期

【摘要】：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(P0.01)。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。
[Abstract]:Objective: To investigate the effect of the expression of epidermal growth factor like domain protein 6 (epidermal growth factorlike-domain 6, EGFL 6) in human melanoma A375 cells by specific small interference RNA (small interference RNA, Si RNA), and to explore the effect of EGFL 6 gene silencing on the proliferation and invasion of human melanoma cells. Methods: the target 6 gene was designed and synthesized. The siRNA (named EGFL6-siRNA) was transfected into the melanoma A375 cells by liposome transfection, and the expression of EGFL6 mRNA and protein in melanoma A375 cells was detected by real-time fluorescence quantitative PCR and Western blotting. The cell count kit -8 (cell counting kit-8) method and the experiment were respectively used. The changes in the proliferation and invasion ability of A375 cells were detected. Results: after successful transfection of EGFL6-siRNA 48 h, the expression level of EGFL6 mRNA and protein in A375 cells decreased significantly (P value was 0.01). The proliferation ability of A375 cells in.EGFL6-siRNA transfected group was significantly inhibited (P values were 0.01) at 24~72 H (0.01), and cell invasiveness Conclusion: P0.01. Conclusion: specific siRNA can effectively silence the expression of EGFL 6 gene in human melanoma A375 cells and inhibit the proliferation and invasion of tumor cells.

【作者单位】：广东省分子免疫与抗体工程重点实验室暨南大学抗体工程研究中心;
【基金】：国家自然科学基金资助项目(编号:81372281)~~
【分类号】：R739.5

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