Figure 4. Cx32 suppresses HCC cell proliferation through inhibition of the Akt signaling pathway.

(A, B) EdU assay analysis of the Cx32 knockdown (A) and overexpression (B) effect on the proliferation of HepG2 and SMMC-7721 cells. Cells were cultured in 24-well microtiter plates after transfection or non-transfection. EdU (100 μM) was added, and the cells were cultured for another 2 h before EdU and Hoechst staining. The total cell number and EdU-positive cell number were counted in five random fields; ** p < 0.01. (C) The expression of PCNA, Akt, and cell cycle regulatory proteins in Cx32-overexpression or knockdown HCC cells and control cells. (D) The PI3K inhibitor attenuated the inhibitory function of Cx32 on PCNA and cyclin D1 protein levels, as examined by western blotting. Data were measured as relative density (RD) of p-Akt/Akt, Cyclin D1/Actin, and PCNA/Actin. The first control lane was defined as 1.