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目的探讨抑制COMMD7基因后对肝癌干细胞(liver cancer stem cells,LCSCs)侵袭转移能力的影响及其机制。方法使用shRNA慢病毒转染抑制COMMD7在LCSCs中的表达,粘附、Transwell实验检测细胞侵袭迁移能力,鬼笔环肽染色检测细胞形态学特征,Western blot检测细胞中E-cadherin、N-cadherin、Vimentin蛋白表达。结果抑制COMMD7基因表达可以显著降低LCSCs的侵袭迁移能力,对照组与转染组粘附相对细胞量为(1.00±0.12),(2.35±0.20),转染组粘附细胞数量显著增加(P<0.05)。对照组与转染组每视野迁移相对细胞量为(1.00±0.04),(0.24±0.03),每视野侵袭相对细胞量为(1.00±0.05),(0.24±0.04),转染组迁移侵袭细胞数明显降低(P<0.05),同时导致细胞表现出低侵袭迁移能力的形态学变化,上述改变伴随了间充质-上皮转化(mesenchymal-epithelial transition,MET)相关分子E-cadherin表达上升(P<0.05),N-cadherin、Vimentin表达下调(P<0.05)。结论 COMMD7可能通过调控MET过程抑制LCSCs的侵袭转移能力。
Objective To investigate the effect of COMMD7 gene on invasion and metastasis of liver cancer stem cells (LCSCs) and its mechanism. Methods The expression of COMMD7 in LCSCs was inhibited by shRNA lentiviral transfection. The invasion and migration of COMMD7 were detected by Transwell assay. The morphological features of cells were detected by phalloidin staining. The expressions of E-cadherin, N-cadherin, Vimentin protein expression. Results Inhibition of COMMD7 gene expression could significantly reduce the invasion and migration of LCSCs. The relative cell numbers of the control group and the transfected group were (1.00 ± 0.12) and (2.35 ± 0.20), respectively. The number of adherent cells in the transfected group was significantly increased (P < 0.05). The relative amount of cells per field of view in the control group and the transfection group was (1.00 ± 0.04), (0.24 ± 0.03), and the relative cell number per field of view was (1.00 ± 0.05), (0.24 ± 0.04) (P <0.05), and resulted in the morphological changes of cells with low invasion and migration. The above changes accompanied with the increase of E-cadherin expression in mesenchymal-epithelial transition (P <0.05). The expressions of N-cadherin and Vimentin were down-regulated (P <0.05). Conclusion COMMD7 may inhibit the invasion and metastasis of LCSCs through regulating the MET process.