Fig. 4

Up-regulation of Gadd45β by 5-azacytidine increased the chemosensitivity of liver cancer cells through decreasing the stemness of HCC cells. a After SMMC-7721 and Hep-3B cells were treated with 5-azacytidine, the methylation-specific PCR (MSP) assay showed that methylation of Gadd45β promoter was obviously inhibited. b Western blot assay confirmed that the cells treated with 5-azacytidine promoted the expression of Gadd45β protein. c, d RT PCR then indicated that 5-azacytidine inhibited the mRNA expression of SOX9 and OCT4 in SMMC-7721 and Hep-3B cells. GAPDH was used as an internal reference (*P < 0.05; **P < 0.01). e Flow cytometry was then used to detect the apoptosis of cells. the result showed that 5-azacytidine treatment significantly enhanced the cisplatin-induced the apoptosis of SMMC-7721 and Hep-3B cells than control group. f, g Quantification of the apoptosis of cancer cells. Data are presented as the mean ± SD (**P < 0.01)