The Effect Of Thiomyristoyl On The Occurrence And Development Of Leukemia Cells And Its Relationship With Sirt2

Thiomyristoyl?sulfated myristoyl lysine compound,TM?is a novel inhibitor that specifically inhibits Sirt2,a NAD⁺?nicotinamide adenine dinucleotide?-dependent deacetylase.Sirt2 can deacetylate a variety of acetylated proteins and can be widely involved in cellular activities.In recent years,it has been found that Sirt2 is highly expressed in various tumors.so it is often used as a therapeutic target for tumors.Among them,more studies have shown that Sirt2 is highly expressed in AML leukemia patients,and the expression level is higher in relapsed patients.At the same time,Sirt2 is also highly expressed in human primary MLL-r AML leukemia cells.It is suggested that Sirt2 may play an important role in the development of MLL-r AML leukemia,and it can be used as a therapeutic target for this type of leukemia.TM as a highly selective inhibitor of Sirt2 may also be a potential drug for leukemia treatment.The aim of this study was to investigate the role of TM in the development of MLL-r AML leukemia and its relationship with Sirt2 gene,and to explore the molecular mechanism and therapeutic regimen of its action,providing a preliminary theoretical and experimental basis for the treatment of MLL-r AML leukemia with TM.In this study,first,we used TM to treat human leukemia cells,and detected the expression level of intracellular Sirt2 substrate protein H4K16Ac by Western blot,and compared the treated cells with cell functional experiments.Including the detection of clone formation ability,the detection of cell viability by MTT method,the detection of cell cycle and apoptosis by flow cytometry.Subsequently,in order to study the relationship between TM action and Sirt2 gene expression,we first constructed the Sirt2 knockdown cell line by shRNA interference technology,and used MTT assay to detect the effect of TM on Sirt2 knockdown cell viability.The mouse-derived MLL-AF9,MLL-ENL,sirt2^-/-MLL-AF9 and sirt2^-/-MLL-ENL leukemia cell lines were further selected and compared by related cell function tests.Then,in order to study the role of TM in the vivo of mouse,we conducted related experimental studies through different doses,administration methods,transplant cell types and methods,and so on;Finally,to investigate whether the molecular mechanism of TM action is related to c-Myc protein,we examined the effect of TM treatment on the expression of c-Myc protein in murine MLL-r cells by Western blot.The results of this thesis show that TM can inhibit the growth of human AML leukemia cells and reduce the expression of H4K16Ac protein.At the same time,TM can cause cell cycle arrest in G1 phase and promote cell apoptosis.However,when studying the relationship between TM action and Sirt2 gene expression,MTT assay showed that the effect of TM treatment on Sirt2 knockdown cell line was not obvious compared with the control group;the results of various cell functional experiments on mouse-derived cells found that TM pair Sirt2-/-MLL-r cells have the same inhibitory effect as MLL-r cells.This indicates that the inhibitory effect of TM on MLL-r AML leukemia cells is not related to Sirt2 gene expression.Several results of TM experiments in transplanted mice found that for direct in vivo administration of TM,drug toxicity caused a decrease in survival rate in mice,and in vivo experiments on TM have yet to be further studied.In the results of testing the effect of TM on c-Myc protein,it was found that when murine MLL-r cells no longer expressed Sirt2 gene,the expression of c-Myc protein was significantly reduced,while the expression level of c-Myc protein in the cells could be increased by TM treatment.When cells normally express Sirt2 gene,TM treatment can reduce the expression level of c-Myc protein in cells.Unexpectedly,the experiment detected that the expression level of c-Myc short chain protein was reduced by TM treatment.These results indicate that the inhibitory effect of TM on MLL-r AML leukemia cells is related to the change of c-Myc expression,and it is related to whether the cells normally express Sirt2 gene,but the specific mechanism remains to be further studied.Therefore,this paper demonstrates the inhibitory effect of TM on AML leukemia cells,and conducts preliminary research on the related molecular mechanisms.The mouse pre-experiment provides a reference strategy for studying treatment of leukemia by TM in vivo.These findings have important implications for the use of TM in the treatment of MLL-r AML leukemia.