Fig. 6

Increased E-cadherin expression in HCC treated with 5-AZA and vitamin C. a Quantitative changes in E-cadherin mRNA in HLE and Huh7 treated with vitamin C, 5-AZA, and 5-AZA + vitamin C compared to the untreated control after 48 h. Data was normalized using GAPDH expression as a reference control. b Western blot analysis for changes in P21 and cyclin B1 proteins in HLE and Huh7 with various treatments. Calculations were normalized with GAPDH protein. All the data are the average of the experiments (N = 3). Statistical significance was tested using one-way ANOVA (non-parametric) Tukey’s test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.01). Error bars represent the standard deviation. c Vitamin C enhances active demethylation induced by 5-AZA, in converting 5-methyl cytosine to 5-hydroxymethyl cytosine by increasing the expression of TET2 and TET3. Induced demethylation leads to decrease in Snail expression and upregulation of GADD45B. Reduced Snail expression leads to upregulation of P21. The network of interactions between GADD45, P21, cyclin B1, and PCNA leads to the arrest of cells in G1, S, or G2/M phases of the cell cycle. On the other hand, increased expression of E-cadherin by reduction of Snail leads to the inhibition of EMT