南方医科大学学报2018，Vol.38Issue(5)：527-533,7.
全反式维甲酸诱导肝癌细胞Hepa1-6成熟和分化过程中的自噬
Changes in autophagy during maturation and differentiation of Hepa1-6 cells induced by all-trans retinoic acid
摘要
Abstract
Objective To investigate the effects of different concentrations of all-trans retinoic acid (ATRA) on the maturation, differentiation and autophagy of Hepa1-6 cells.Method Hepa1-6 cells were treated with 0.1,1,and 10 μmol/L ATRA,and the changes in the expressions of hepatic specific markers were detected using real-time PCR and Western blotting.Indocyanine green(ICG)and periodic acid-schiff(PAS)staining was used to assess the functional maturation of Hepa1-6 cells,and the cell-cell junction and autophagy were observed under transmission electron microscopy to determine the optimal concentration of ATRA for treatment. The expressions of autophagy-related markers in the cells were detected using Western blotting, and confocal microscopy was used to observe the autophagic flow in the cells transfected with ptfLC3 plasmid.Results Compared with the control cells,the hepatocytes treated with ATRA showed a concentration-dependent decrease in AFP expression and increase in the expressions of ALB, CK18, TAT and ApoB. ICG and PAS staining revealed significantly increased number of positive cells after ATRA treatment. Following ATRA treatment, the cells exhibited obviously increased tight junctions, cytoskeleton and number of autophagosomes under transmission electron microscopy. ATRA treatment resulted in significantly increased the expressions of autophagy-related markers LC3-II, Beclin-1, RAB7 and P62 and also an increased ratio of LC3-Ⅱ/LC3-Ⅰ(P<0.05).Confocal microscopy revealed obviously increased green and red spots in the cells after ATRA treatment. Conclusion ATRA can induce the maturation and differentiation and enhance the level of autophagy in Hepa1-6 cells.关键词
原发性肝癌/全反式维甲酸/自噬/细胞分化Key words
primary hepatic carcinoma/all-trans retinoic acid/autophagy/cell differentiation引用本文复制引用
方姝煜,崔洁洁,龚梦嘉,何昀,张敬芳,毕杨..全反式维甲酸诱导肝癌细胞Hepa1-6成熟和分化过程中的自噬[J].南方医科大学学报,2018,38(5):527-533,7.基金项目
重庆市自然科学基金(csct2016jcyjA0228) （csct2016jcyjA0228）
