Anti-tumor Effects And Mechanisms Of Cell Cycle Protein WEE1 In Acute T-cell Lymphoblastic Leukemia

Objective: Acute T lymphocytic leukemia(ALL)is a kind of malignant tumor which originated from the abnormal proliferation of T lymphocytes in bone marrow.Malignant tumor originated from the abnormal proliferation of T lymphocytes in bone marrow.However,the pathogenesis of acute T lymphocytic leukemia is complicated,and the recurrence rate is high,but most of them are very difficult to cure.In this study,we investigate the mechanism of MK1775,an inhibitor of WEE1,which is related to cell cycle regulation.And the treatment of T-ALL with the combination of transglutaminase GLS1 inhibitor CB839,which is expected to provide a new method for clinical treatment in the late stage of T-ALL.Methods: In this study,we examined the expression level of WEE1 by western blot analysis in a variety of T-ALL cell lines.CCK8 is used to assay the activity of T-ALL cells which are treated with MK1775.Lentiviral infection is used to silencing WEE1 in T-ALL cells,after the infection we count the cell number at 1,3,5,7 days to observe the effect of cell growth.Through the GSE analysis,we find c-MYC combines on the transcription start union of WEE1.The UCSC data also show above result.Then we use chromatin immune coprecipitation experiment to prove it.Additionally,cell apoptosis were assessed by FACS to observe the combination of MK1775 and CB839.Results: Oncomine data analysis results showed that patients with T-ALL express higher WEE1,which compared with nomal.And western bloting verified that WEE1 protein lever was significantly higher than that of nomal cells.CCK8 assay was used to dectect the sensitivity of T-ALL cell line to WEE1 inhibitor MK1775,and the majority of T-ALL cell lines' IC50 was below 400 nM.At the same dose MK1775 hasstrong effect on T-ALL cells,but,it is almost non-toxic to nomal cells.The silencing of WEE1 in KOPT-K1 and CCRF-CEM was slowed the growth of cells,indicating that WEE1 plays an important role in the proliferation of T-ALL.UCSC data analysis showed that c-MYC and WEE1 had a certain correlation in T-ALL,we verified that c-MYC did regulate the transcription of WEE1 by ChIP experiment.And,we find CB839 and MK1775 show good synergistic effect in T-ALL cell line.Conclusion: Our research showed that WEE1 was highly expressed in T-ALL,at the same time T-ALL cell lines showed high sensitivity to the WEE1 inhibitor MK1775.Though big data analysis and chromatin immune coprecipitation experiment,we find c-MYC directly combine on WEE1 and activate its transcription.In view of the single targeted therapy is easy to produce drug resistance,through the experiment we find the combination of MK1775 and CB839 showed a good synergistic effect,which provid a new way for the treatment of T-ALL.