All-trans retinoic acid reverses malignant biological behavior of hepatocarcinoma cells by regulating miR-200 family members

As a potential chemo-therapeutic agent,all-trans retinoic acid(ATRA)can signif-icantly reverse epithelial-mesenchymal transition(EMT)of hepal-6 hepatocarcinoma cell line in vitro,but the mechanism is unclear.The expression profile of microRNA-200(miR-200)families is different in hepatocellular carcinoma.In this study,we found that ATRA trea tment could up-regulate the expression of miR-200a-3p,200c-3p,and 141-3p,which were involved in ATRA regulated proliferation and apoptosis of hepal-6 cell,but not colony formation.Meanwhile,miR-200a-3p,200c-3p,and 141-3p could recovery ATRA inhibited migration and invasion abilities of hepal-6 cells at various levels.miR-200a-3p and 200c-3p prevented ATRA from inducing the differentia tion and hepatic functions of hepal-6 cells.An-tagomir specific for miR-200a-3p and 200c-3p down-regulated the expression of CK18,but only miR-200a-3p anta gomir played prominent role in regula ting the expression of these mesenchymal markers,N-Cadherin,Snail and Twist.The transcriptional activities of 8 tran-scription factors were up-regulated and 35 transcription factors were down-regulated by ATRA.Compared with ATRA group,inhibition of miR-200a-3p,200c-3p,and 141-3p significantly strengthened the expression of Fra1/Jun(AP1),Ets1/PEA3,Brn3,and Zeb1/AREB6 at varying degrees.Therefore,this result suggested that ATRA may suppress EMT through down-regulating miR-200a-3p,200c-3p and 141-3p related transcription factors.miR-200 and their downstream genes might be the potentially specific targets for the treat-ment of hepatocarcinoma.