Effect of ADAM10 inhibitor GI254023X on proliferation and apoptosis of acute T-lymphoblastic leukemia CCRF-CEM and Molt4 cells and its possible mechanisms

Objective To investigate the effect of GI254023X, an ADAM10 (a desintegrin and metalloproteinase domain containing protein 10)inhibitor, on the proliferation and apoptosis of CCRF-CEM and Molt4 cell lines and its mechanisms. Methods CCRF-CEM and Molt4 cells were treated with different concentrations of GI254023X. Proliferation-inhibition curve was assayed and plotted by CCK-8 method. Cell viability and apoptosis were detected by flow cytometry with Annexin V and 7-AAD staining, and cell cycle changes were analyzed by flow cytometry. For mechanism exploration, the cleavage of Notch1 protein was determined by Western blot. The transcripts 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 rvalues of 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). The half maximal inhibitory concentration (IC₅₀) values of CCRF-CEM cells for 24 and 48 h were (29.40±3.28) μmol/L and (11.14±2.21) μmol/L, respectively, while IC₅₀ values of Molt4 cells for 24 and 48 h were (19.04±1.58) μmol/L and (11.25±2.31) μmol/L, respectively. Compared with the control group,the apoptosis rate of cells increased with the increment of concentration of GI254023X. Cell cycles were arrested at G₀/G₁ phase. Compared with DMSO in the control group, the expression 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 Notch1 target gene Hes-1 mRNA transcripts in CCRF-CEM and Molt4 cells were reduced in GI254023X treated group, while the mRNA levels of pro-apoptosis genes BAD, BAK increased. Conclusions GI254023X can inhibit the proliferation and induce apoptosis of CCRF-CEM and Molt4 cells.Its mechanism may be related to the inhibition of Notch1 pathway.