摘要
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 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 ceils 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-eadherin and Vimentin（ P 〈 0.05 ） , the molecules related knockdown inhibits the invasion and course to mesenchymal-epithelial transition migration in LCSCs, which may be （MET）. Conclusion through its regulation COMMD7 on the MET
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2017年第17期1691-1695,共5页
Journal of Third Military Medical University
基金
国家自然科学基金青年科学基金(81402032)
国家自然科学基金面上项目(81672902)~~
关键词
COMMD7
肝癌干细胞
侵袭转移
COMMD7
liver cancer stem cells
invasion and migration
