Abstract:
ob<x>jective: To investigate the effects of ADAM10 inhibitor GI254023X on the proliferation and apoptosis of acute T-lymphoblastic leukemia CCRF-CEM and Molt4 Cells and its mechanisms.Methods: CCRF-CEM and Molt4 Cells were treated with different concentrations of GI254023X. Proliferation-inhibition Acurve was assayed and plotted by CCK-8 method.cell viability and apoptosis was detected by flow cytometry with Annexin V and 7-AAD staining.Cell cycle changes were analyzed by flow cytometry.The cleavage of Notch1 protein was determined by Western blot.The transc<x>ripts of anti-apoptotic genes Bcl-2、c-Myc、Mcl-1 ,pro-apoptosis genes BAD、BAK and Notch1 target gene Hes-1 were detected by Real-time PCR .Results: GI254023X inhibited the proliferation of CCRF-CEM and Molt4 Cells in a time-and dose-depended manner(the r valuesof CCRF-CEM cells for 24 and 48 h were 0.981 and 0.962 respectively, while r values of Molt4 Cells for 24 and 48 h were 0.992 and 0.957 respectively. ), and the IC50 values of CCRF-CEM cells for 24 and 48 h were (29.403±3.28)μmol/L and (11.14±2.21)μmol/L respectively, while IC 50 values of Molt4 Cells for 24 and 48 h were (19.042±1.58)μmol/L and (11.25±2.31)μmol/L respectively. As compared with the control group ,the apoptosis of cells increased with concentration of GI254023X; Cell cycles were arrested at the G0/G1 phase.Compared with DMSO in control group,The ex<x>pression of Cleaved Notch1 was down-regulated after the treatment with GI254023X .The mRNA levels of anti-apoptotic genes Bcl-2、c-Myc、Mcl-1 and Hes-1 mRNA transc<x>ripts in CCRF-CEM and Molt4 Cells were reduced in GI254023X treated group, and the mRNA levels of pro-apoptosis genes BAD、BAK were increased. Conclusion: GI254023X can remarkably inhibit cell proliferation and induce apoptosis in CCRF-CEM and Molt4 Cells.Its mechanism may be associated with inhibition of Notch1 activation