Objective To study whether emodin combined with 5-AzA-2'-deoxycytidine(5AzA-cdR)can enhance its demethylation effect on a tumor suppressor gene ppENK of pancreatic cancer cells.Methods Panc1 cells were se-lected as the research object.First,cell count test(CCK-8)was used to detect the growth inhibition of different concen-trations of emodin on pancreatic cancer cells.According to the results of CCK-8,the optimal concentration of emodin was selected.Later,the Panc1 cells were divided into four groups:control group,emodin group,5AzA-cdR group,and emodin combined with 5AzA-cdR group.Next,Dot-blot was used to detect the effects of four groups on genomic 5-methylcyto-sine(5mc),and finally,bisulfite sequencing PCR(BSP)was used to detect the effects of four groups on ppENK methyl-ation.Results CCK-8 shows that emodin can inhibit the growth of Panc1 cells through concentration and time gradi-ents.The Dot-blot results showed that the 5mc levels in the emodin group and 5AzA-cdR group were lower than those in the control group,and the differences were statistically significant(t=12.55,20.12,P＜0.05).The 5mc level in the 5AzA-cdR group was significantly lower than that in the emodin group,and the difference was statistically significant(t=17.76,P＜0.05).The emodin com-bined with 5AzA-cdR group showed a significant decrease in 5mc level compared to the control group,emodin group,and 5AzA-cdR group,and the difference was statistically significant(t=30.22,20.77,10.45,P＜0.05).The BSP results showed that the average level of ppENK methylation in the emodin group and 5AzA-cdR group was lower than that in the control group,with statistical significance(t=14.27,21.34,P＜0.05).The methylation level in the 5AzA-cdR group was significantly lower than that in the larger emodin group,with statistical significance(t=16.41,P＜0.05).The group treated with emodin combined with 5AzA-cdR showed a significant decrease in methylation level compared to the control group,emodin group,and 5AzA-cdR group,and the differences were statistically significant(t=32.68,19.33,10.12,P＜0.05).Conclusion Emodin com-bined with 5AzA-cdR can enhance the demethylation of tumor suppressor gene ppENK in pancreatic cancer cells.
维普
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