Study On The Apoptosis Effect And Its Mechanisms Of Leukemia MOLT-4 Cells Induced By Resveratrol

Objective : To study the inductive effect on the growth,induce-cell apoptosis activity and its mechanisms of resveratrol to lymphoblast leukaemia Molt-4 cells in vitro. To provide the experimental evidence for the clinical application of resveratrol on leukaemia.Method:1 The aim of study was human lymphoblast leukaemia MOLT-4 cells.Control group and treated group with different concentrations resveratrol (final concentration :12.5,25,50,100,200μmol/L)was setted .MTT method was used to detect the suppressing effect of resveratrol on human leukaemia MOLT-4 cells for 24h,48h and 72h.2 Control group and treated group was settled.Wrigh-Giemssa,transmission electron microscope technique were used to detect the apoptosis status of MOLT-4 cells treated with different concentrations resveratrol for 48 hours.3 Flow cytometry was performed to observe cell cycle distribution and apoptosis percentage of control group and treated group with different concentrationsresveratrol(50,100μmol/L)on MOLT-4 cells for 48 hours.4 MOLT-4 cells were treated with different concentrations(0, 50, 100μmol/L) of resveratrol for 48 hours .The RT-PCR was performed to detect the levels of WAVE1mRNA, to explore the induced-cell apoptosis activity and its mechanisms of resveratrol to MOLT-4 cells.Results:1 Compared with control group, resveratrol could inhibit the proliferation of MOLT-4 cells (p<0.05) after treated with different concentrations resveratrol for 24,48,72hours,the inhibition ratio as follows:12.5μmol/L resveratrol was 3.84%,17.52%,29.32%; 25μmol/L resveratrol was 8.77%,23.86%,36.11%;50μmol/L resveratrol was 11.38%,32.79%,53.92%; 100μmol/Lresveratrol was 19.24%,43.33%,62.50% ; 200μmol/L was 26.51%,57.64%,74.98%,the most inhibitionary 74.98% was treated with 200μmol/L resveratrol for 72hours.Under the same time,the inhibitory effect was enhanced with the concentrations,Under the same concentration resveratrol,the inhibitory effect was enhanced with the time.Resveratrol inhibited the proliferation of MOLT-4 cells in a time-and-dose-dependent manner.2 Compared with control group,after treated with resveratrol (100μmol/L)for 48 hours,the MOLT-4 cells shown some typical morphologic feature and super-microstructure changes of apoptosis,including cell shrinkage,chromatin concentration,karyotheca forming outwardly acute angle prominency,cytoplasm vacuolar degeneration,mitochondrion and rough endoplasmic reticulum reduction,karyopyknosis and karyorrhexis.3 Compared with control group,after MOLT-4 cells were treated with different concentrations resveratrol for 48 hours,the rates of G0/G1 and G2/M phase cells were distinctly declined,and the rate of S phase cells were increased from (42.2±0.73)%to(78.1±0.62)%,resveratrol could arrest cell cycle in S phase;The apoptosis rate of control geoup was(1.52±0.09)%,but the apoptosis rates of treated with resveratrol were : 50μmol/L(7.11±0.30)%,100μmol/L(17.8±0.40)%.4 After treated with different concentrations resveratrol (50,100μmol/L) for 48 hours. RT-PCR technique indicated,the mRNA express adjusted values of WAVE1/GAPDH in untreated cells were 0.586±0.06, after treated with 50μmol/L and 100μmol/L resveratrol ,the mRNA express adjusted values of WAVE1/GAPDH were 50μmol/L 0.356±0.03;100μmol/L 0.382±0.05.Compared with control group,after treated with resveratrol,the mRNA expression of WAVE1 was reduced.Conclusions : Resveratrol can inhibit the proliferation of leukaemia MOLT-4 cells in a time-and-dose-dependent manner,Resveratrol can arrest cell cycle and induce apoptosis in MOLT-4 cells,and the cells were biocked in S phase;which may be associated with downregulating the expression of WAVE1 to promote apoptosis.