Study On The Expression Of Htert Gene Mrna And Promoter Methylation Status Of Htert In Human Leukemia Hl-60 Cell Line By Using Cantharidin

Objectives:To investigate the expression of hTERT gene mRNA and promoter methylation status of hTERT in human leukemia HL-60 cell line by using cantharidin and indicate if cantharidin' mechanism of anti-leukemia is related with its regulation of hTERT promoter methylation level to regulating hTERT expression.Methods:Using MTT method to confirmed the exact role of cantharidin inhibit proliferation of HL60 cell line; Using inverted microscope to observe the cell morphology after the drug and to take a photograph. To determine the mRNA expressions of hTERT in HL60 cell line after using different concentrations of cantharidin by RT-PCR assay; To determine the promoter methylation status of hTERT in HL60 cell line after using different concentrations of cantharidin by MSP assay.Results:Cantharidin inhibit the proliferation of HL60 cells with obvious time-and dose effects;The mRNA relative expressions of hTERT are significantly different among Control group,Moderate doses cantharidin group and High doses cantharidin group(P<0.01);Moderate doses cantharidin group and High doses cantharidin group can lower the mRNA relative expressions of hTERT(P<0.01); The mRNA relative expressions of hTERT in High doses cantharidin group are lower than Moderate doses cantharidin group(P<0.01);The hTERT promoter are in Methylation status in all the 3 groups. Conclusion:The effect of cantharidin' inhibiting HL60 cell proliferation is related with its lowering the mRNA expressions of hTERT; The cantharidin' regulation of hTERT promoter methylation level need further research by Real-time quantitative MSP assay to confirm.