The Effects And Mechanisms Of Resveratrol On RUNX3in Murine Melanoma

Part1:The Effects of Resveratrol on RUNX3gene in B16F10Murine Melanoma cellObjectives:To explore the effects and mechanism of RES on RUNX3gene in B16F10cell line. Methods:The B16F10cell line were treated with DMSO、different concentration of RES(20μM,40μM,80μM) and positive drug [(5-Aza-CdR),10μM]for48h. The difference of mRNA expression and protein expression and the promoter region methylation status of the RUNX3gene in B16F10cell were measured by using RT-PCR, Western-Blotting and MSP, respectively. RESULT:1) No expression of RUNX3mRNA and protein were found in untreated B16F10cell. After the B16F10cell treated by methylation inhibitior5-Aza-CdR, expression of RUNX3mRNA and protein were found.2) After the B16F10cell treated by different concentration of RES, expression of RUNX3mRNA and protein were found.And the degree of the expression was directly coorelated with concerntration of RES.3) RUNX3gene promoter was hypermethylated in untreated B16F10cell.After treated by RES, the promoter of RUNX3gene was demethylated partly and is coorelated with the concentration of RES.PAR2:The Effects of Resveratrol on RUNX3gene in C57BL mice bearing B16F10melanomaObjectives:To evaluate the effects of RES on RUNX3gene in C57BL mice bearing B16F10melanoma. Methods:B16F10melanoma cells were injected subcutaneously in C57BL mice.And C57BL mice bearing B16F10melanoma were treated with10%CMC-Na, RES(50、100mg/kg/d), positive drug(DTIC40mg/kg/3d) for21d.At the last day,the C57BL mouse were sacrificed.Tumor growth was assessed by direct measuring weight and volume of the solid tumor.Levels of RUNX3mRNA in peripheral blood and tumor tussie were determined by RT-PCR,and expression of RUNX3protein was analysed by Western-Blotting. Result:1) In vivo, low dose RES, hight dose RES and DTIC could inhibite the growth of tumor in C57BL mice, and the tumor volume was decreased to0.83-fold,0.59-fold and0.27-fold respectively (P<0.05or0.01) as compared with control group.2) While no significant changes were detected on expression of RUNX3mRNA and protein in DTIC group, the expression of RUNX3mRNA and protein in RES group was significantly increased.Conclusion:RES can increase the expression of RUNX3in melanoma B16F10cells and C57BL mice bearing B16F10melanoma; Suggesting that one of the potential mechanisms may be related with Demethylation of RES.