The Effect Of Combining Hypomethylating Agents With Histone Deacetylase Inhibitors To Affect THP-1cells

Objective: To observe the effect of demethylating agents combined with histone deacetylase inhibitors on the expression of tumor suppressor gene PTPN6,and to investigate whether the combination of the two drugs can enhance the anti-leukemia effect through the study of leukemia cell lines.Methods:1.Methylation specific PCR(MSP)was used to detect the methylation status of PTPN6 gene in THP-1 cell line.2.Different concentrations of DNMTi(5-azacitidine)and HDACi(chidamide,panobinostat)were used alone or in combination in cell lines,and the changes of PTPN6 m RNA expression were detected by quantitative real-time polymerase chain reaction(qt-PCR),and the changes of PTPN6 protein levels were detected by Western blot.Results: 1.The PTPN6 gene is methylated in the DNA of THP-1 cell line.2.5-azacitidine could affect the expression of PTPN6 m RNA in THP-1 cell lines in a dose and time dependent manner(P<0.05),while the effects of cidabenamine and panobinostat on PTPN6 m RNA were time-dependent(P<0.05).3.When 5-azacitidine reached a certain concentration,DNMTi combined with HDACi significantly increased PTPN6 expression,which was superior to the two alone,and the combination effect was greater than the sum of the effects of the two drugs alone(P<0.05).Conclusion:1.When a certain concentration was reached,5-azacitidine was able to increase the expression level of tumor suppressor gene PTPN6 in THP-1 cells and inhibit the proliferation of tumor cell lines.2.The combination of DNMTi and HDACi can significantly increase the expression of PTPN6 in THP-1 cells,and its effect is superior to the sum of the effects of the two alone.