COMMD7调控肝癌干细胞间充质-上皮转化过程抑制其侵袭转移能力

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.

COMMD7 inhibits migration and invasion in liver cancer stem cells via regulating mesenchymal-epithelial transition

Objective To determine the effect of COMMD7 inhibition on invasion and migration in liver cancer stem cells (LCSCs),and investigate the possible mechanism.Methods After LCSCs were infected by shRNA lentiviral vectors of COMMD7,adhesion assay and Transwell assay were used to detect the invasion and migration,and phalloidin staining was employed to observe the morphological changes.Western blotting was adopted to measure the expression of E-cadherin,N-cadherin and Vimentin.Results COMMD7 knockdown significantly inhibited the invasion and migration of LCSCs.The relative cell quantity of adhesion was 1.00 ± 0.12 and 2.35 ± 0.20 respectively in control cells and infected cells,suggesting there were significantly more adhesive cells in the infected group (P ＜ 0.05).The relative cell quantity per visual field of migration was 1.00 ±0.04 and 0.24±0.03,and that of invasion was 1.00 ±0.05 and 0.24 ±0.04 respectively in the control cells and infected cells,and there were significantly less invasive and migrated cells in the infected group (P ＜0.05).What's more,COMMD7 knockdown also induced some morphological changes of cells corresponding to the weakened abilities of migration and invasion.All the changes above were associated with up-regulation of E-cadherin (P ＜ 0.05) and down-regulation of N-cadherin and Vimentin (P ＜0.05),the molecules related to mesenchymal-epithelial transition (MET).Conclusion COMMD7 knockdown inhibits the invasion and migration in LCSCs,which may be through its regulation on the MET course.