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目的:检测Cullin 4A(CUL4A)表达对卵巢癌细胞迁移及侵袭能力的影响,并初步探讨其相关分子机制。方法:Western blot法检测人卵巢癌细胞株中CUL4A表达,设计合成靶向CUL4A的特异性shRNA,构建稳定低表达CUL4A的卵巢癌细胞株。Western blot和荧光定量PCR验证干扰效率;划痕实验及Transwell法检测下调CUL4A后细胞的迁移及侵袭能力;Western blot和荧光定量PCR检测上皮间质转化(EMT)相关指标及上游信号分子Snail、ZEB1变化。结果:SKOV-3、OV2008、ES-2、OVCAR-3卵巢癌细胞中CUL4A表达均高于正常卵巢上皮细胞HOSE,且以SKOV-3表达水平最高。shRNA转染SKOV-3细胞后,CUL4A的mRNA及蛋白水平显著下降,细胞迁移和侵袭能力明显受到抑制;E-cadherin表达增加,N-cadherin、Vimentin、Snail和ZEB1表达均显著降低。结论:CUL4A可促进卵巢癌细胞的迁移和侵袭,其分子机制可能与调节EMT的上游信号分子Snail及ZEB1有关,提示CUL4A在卵巢癌的发生发展中起着重要的作用。
AIM: To investigate the effect of Cullin 4A (CUL4A) expression on the migration and invasion of ovarian cancer cells and to investigate the underlying molecular mechanisms. Methods: The expression of CUL4A in human ovarian cancer cell lines was detected by Western blot. The specific shRNA targeting CUL4A was designed and synthesized, and the ovarian cancer cell line stably expressing CUL4A was constructed. The interference efficiency was verified by Western blot and quantitative real-time PCR. Scratch assay and Transwell assay were used to detect the migration and invasion ability of CUL4A cells. Western blot and fluorescent quantitative PCR were used to detect the expression of epithelial mesenchymal transition (EMT) and related signaling molecules Snail and ZEB1 Variety. Results: The expression of CUL4A in SKOV-3, OV2008, ES-2 and OVCAR-3 ovarian cancer cells was higher than that in normal ovarian epithelial cells, and the expression level of SKOV-3 was the highest. CUL4A mRNA and protein levels were significantly decreased after SKOV-3 cells were transfected with shRNA, and cell migration and invasion were significantly inhibited. The expression of E-cadherin and the expressions of N-cadherin, Vimentin, Snail and ZEB1 were significantly decreased. CONCLUSION: CUL4A can promote the migration and invasion of ovarian cancer cells. The molecular mechanism may be related to the regulation of EMT upstream signal molecules Snail and ZEB1, suggesting that CUL4A plays an important role in the development of ovarian cancer.