Mechanism Of Action Of Resveratrol On Molt-4Cells Through Notch1Signaling Pathway

Objective:Cell culture technique was employed, to observe the effect ofresveratrol on proliferation and apoptosis of human T-ALL Molt-4cells, andto explore the mechanism of anti-T-ALL effect of resveratrol through Notch1signaling pathway, providing more experimental evidence for the clinicalapplication of resveratrol on leukaemia. We also try to observe the synergisticeffect of DAPT and resveratrol combined with DAPT on proliferation andapoptosis of Molt-4cells, which may have potential clinical value.Methods:1MTT assay were used to detect the inhibition rate of cell proliferationof different concentrations resveratrol（50、75、100、125μmol/L） on humanT-ALL Molt-4cells for24h,48h and72h.2Flow cytometry were used to detect the apoptosis rate of control groupand treated groups with different concentrations resveratrol（50、75、100、125μmol/L） on human T-ALL Molt-4cells for24hours.3RT-PCR were used to detect the expression of Notch1and Hes-1mRNA of control group and treated groups with different concentrationsresveratro（l50、75、100μmol/L） on human T-ALL Molt-4cells for24hours.4MTT assay were used to detect the inhibition rate of cell proliferationof DAPT and different concentrations resveratrol（50、75、100、125μmol/L）combined with DAPT （10μmol/L） on human T-ALL Molt-4cells for24h,48h and72h.5Flow cytometry were used to detect the apoptosis rate of control groupand treated group with DAPT （10μmol/L）and different concentrationsresveratrol（50、75、100、125μmol/L）combined with DAPT （10μmol/L）on human leukaemia on Molt-4cells for24hours. Results:1Resveratrol could inhibit the proliferation of Molt-4cells after treatedwith different concentrations resveratrol for24、48、72hours,the inhibitionratio as follows:50μmol/L resveratrol group was3.84%、17.52%、29.32%;75μmol/L resveratrol group was8.77%、23.86%、36.11%；100μmol/Lresveratrol group was11.38%、32.79%、53.92%；125μmol/Lresveratrol groupwas19.24%、43.33%、62.50%； Under the same time,the inhibitoryeffect was enhanced with the concentrations,Under the same concentrationresveratrol,the inhibitory effect was enhanced with the time.Resveratrolinhibited the proliferation of Molt-4cells in a time-and-dose-dependentmanner.2Compared with control group, resveratrol could inducing apoptosis inMolt-4cells after treated with different concentrations resveratrol for24h.Byflow cytometry, the countrol group apoptosis was （8.23±1.03）%,while thedifferent concentrations resveratrol （50、75、100、125μmol/L） were（17.60±1.68）%、（25.07±0.68）%、（43.30±2.11）%、（56.87±1.60）%。:withincreased concentrations of Res, Molt-4cells apoptosis rate was also increased(P<0.05), in a concentration-dependent manner.3Assume the Notch1relative gene expression quantity of control groupis1,the test groups is0.81±0.01、0.72±0.02、0.49±0.01; Assume the Hes-1relative gene expression quantity of control group is1,the test groups is0.33±0.01、0.30±0.01、0.26±0.01;RT-PCR test results showed that, comparedwith that in control group, gene expression of Notch1and Hes-1in Molt-4cells was significantly decreased in Res groups (P<0.05); increase in Resconcentration led to decreased gene expression levels, and the decreased levelwas concentration-dependent.4DAPT could inhibit the proliferation of Molt-4cells (P<0.05) for24、48、72h, the inhibition ratio as follows:19.53%、25.03%、30.89%. The resultsshowed that in different culture time, namely,24h,48h and72h, inhibition rateof Molt-4cell proliferation was gradually increased (P<0.05); while theinhibition ratio of combined groups as follows: Res (50μmol/L)+DAPT（10μmol/L）after24h,48h,72h:55.95%、69.20%、79.55%;(75μmol/L)+ DAPT （10μmol/L） after24h,48h,72h:62.77%、80.03%、86.45%；(100μmol/L)+DAPT（10μmol/L）after24h,48h,72h:69.94%、85.12%、91.18%；(125μmol/L)+DAPT（10μmol/L）after24h,48h,72h:89.77%、93.28%、96.51%. compared with Res group or DAPT group, Res combinedwith DAPT acted on Molt-4cells, which could significantly lead toenhancement of inhibition of the Molt-4cells proliferation and induction ofapoptosis.5Compared with control group,DAPT could inducing apoptosis inMolt-4cells after treated for24h. By flow cytometry, the countrol groupapoptosis was （8.23±1.03）%,while the DAPT （10μmol/L）group was（12.30±1.40）%. Different concentrations resveratrol（50、75、100、125μmol/L）combined with DAPT （10μmol/L）could inducing apoptosis inMolt-4cells after treated for24h.the apoptosis rate as follows:(30.37±1,20)%、(35.13±1.42)%、(65.27±1.9)%、(78.5±2.43)%. Res combinedwith DAPT acted on Molt-4cells, which could significantly lead toenhancement of inhibition of the Molt-4cells proliferation and induction ofapoptosis.Conclusion:Resveratrol might have an anti-T-ALL effect by inhibitinggene expression of Notch1and Hes-1in Notch1signaling pathway; Rescombined with DAPT had a synergistic anti-T-ALL effect. which may have apotential clinical value.