Figure 7.

ATRA-induced autophagy regulated EMT markers in hepatocarcinoma cells and ATRA-induced autophagy via the Bcl-2/Beclin1 pathway independent of mTOR. A. ATRA suppressed the mRNA expression of the mesenchymal markers including Snail, Vimentin, Twist, and N-cadherin and were reversed by 3-MA and Baf. The expression of E-cadherin was increased after ATRA treatment but no statistical difference was found among the ATRA, 3-MA+ATRA, and Baf+ATRA groups in Hepa1-6 cells. The results were confirmed in at least three batches of independent experiments, and representative results are shown. *P<0.05 vs. control group; #P<0.05 vs. ATRA group; B, C. The protein expression of EMT-related proteins were measured by western blot analysis with β-actin normalization in Hepa1-6 and HepG2 cells; D, G. The results of western blot assay showed that the protein expression of PI3K/Akt/mTOR pathway-related markers have no significant variation after ATRA treatment in Hepa1-6 cells and HepG2 cells; E, H. The protein levels of Bcl-2/Beclin1 pathway-related markers were determined using western blot with β-actin normalization in Hepa1-6 cells and HepG2 cells; F, I. An immunofluorescence assay was performed to analyze the protein levels of Bcl-2 and p-Bcl-2 in Hepa1-6 cells and HepG2 cells. Scale bar = 200 μm.