目的 探讨极光激酶A(AURKA)对葡萄膜黑色素瘤(UVM)C918细胞增殖、转移的影响及其分子机制.方法 使用GEPIA数据库分析AURKA表达与UVM病人生存期的关系;使用Lipofectamine 3000将阴性对照的小干扰RNA和AURKA的小干扰RNA转染到C918细胞中构建AURKA敲低的细胞模型;RT-qPCR和 Western Blot实验分别检测AURKA的mRNA和蛋白表达水平;CCK-8和克隆形成实验检测细胞增殖能力;Transwell实验检测细胞侵袭、迁移能力;Western Blot实验检测AURKA和PLK1的蛋白表达水平;GEPIA数据库分析AURKA和PLK1表达在UVM标本中的相关性.结果 与AURKA低表达的UVM患者相比,AURKA高表达的患者生存期显著降低(P＜0.01);与NC组相比,敲低组AURKA的mRNA和蛋白水平明显降低(P＜0.001);相比于NC组细胞,敲低AURKA显著抑制C918细胞的增殖、转移能力(P＜0.001),且PLK1表达水平明显减少.在UVM标本中,AURKA和PLK1的表达显著正相关(P＜0.001).结论 敲低AURKA的表达可以抑制C918细胞的增殖、转移能力,其机制可能与抑制PLK1基因转录有关,提示AURKA激活PLK1转录促进细胞增殖和转移可能是UVM恶性进展的重要机制之一.
Knockdown of AURKA downregulates the expression of PLK1 and inhibits the proliferation and metastasis of uveal melanoma C918 cells
Objective To investigate the effects of aurora kinase A(AURKA)on the proliferation and metastasis of uveal melanoma(UVM)C918 cells and their underlying molecular mechanisms.Methods The relationship between AURKA expression and survival period of UVM patients was analyzed using GEPIA data-base.C918 cells were transfected with negative control small interfering RNA(siRNA)and AURKA-specific siRNA using Lipofectamine 3000 to establish an AURKA knockdown cell model.RT-qPCR and Western Blot experiments were conducted to detect the mRNA and protein expression levels of AURKA,respectively.CCK-8 and colony formation assays were used to evaluate the cell proliferation capacity.Transwell assays were car-ried out to examine the cell invasion and migration abilities.The protein expression levels of AURKA and PLK1 were detected by Western Blot.The correlation between AURKA and PLK1 expression in UVM speci-mens was analyzed by GEPIA database.Results Patients with high AURKA expression exhibited signifi-cantly shorter survival compared to those with low AURKA expression in UVM(P＜0.01).The mRNA and protein levels of AURKA were significantly reduced in the knockdown group compared to the negative control(NC)group(P＜0.001).Knockdown of AURKA significantly inhibited the proliferation and metastasis abili-ties of C918 cells compared to the NC group(P＜0.001),accompanied by a significant decrease in the expres-sion level of PLK1.A significant positive correlation was observed between AURKA and PLK1 expression in UVM specimens(P＜0.001).Conclusion Knockdown of AURKA expression inhibits the proliferation and metastasis abilities of C918 cells,potentially through the inhibition of PLK1 gene transcription.This suggests that AURKA activation of PLK1 transcription to promote cell proliferation and metastasis may be one of the important mechanisms underlying the malignant progression of UVM.
万方数据
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