Study On The Role Of MOF-MSL Complex In Tumorigenesis And Development Of Colon Cancer

Colon cancer is a relatively common malignant disease which is originating from the epithelium of the colonic mucosa,and its incidence is the third among all gastrointestinal tumors.Therefore,early detection and early treatment play indispensable roles in the aspect of the cure of colon cancer patients.In recent years,there has been an increasingly number of research report which proves that the epigenetic regulation mechanism is closely relevant with the tumorigenesis and development of tumors.MOF(males absent on the first)is a member of the histone acetyltransferase MYST family.It acts as a catalytic subunit in cells and participates in at least two multiprotein complexes-MSL and NSL complexes.Although it has been reported that the MOF-MSL complex plays a crucial role in Drosophila’s dose compensation mechanism of the male X chromosome,the exact functions of the two complexes in human cells are unclear.Previous studies have found that the expression of MOF and abnormal level of H4K16 ac modified by MOF are associated with the pathogenesis of a variety of tumors.However,it is unclear whether MOF acts on the pathogenesis of tumors in a free way or in a complex way when tumors occur.In particular,which complex of MOF is related to which kind of tumor remains to be further studied.This article intends to explore the effect of MSL complex on colon cancer by studying the effect of the key subunit MSL1.In this study,the plvx-zsgreen-sh MSL1 gene interference plasmid of the lentiviral system is successfully constructed and 293 T cells is used to package the virus and collect the virus fluid.We know the optimal infection titer and time summarily by observing the expression of green fluorescent protein level through a fluorescence microscope.Then the MSL1 expression is detected at the protein level and m RNA level to determine whether the lentivirus interference MSL1 system is successful.A series of experimental methods such as CCK-8,colony formation,Hoechst staining,Western Blot and RT-PCR are used to explore the effect of MSL1 on the proliferation,apoptosis,invasion and metastasis of colon cancer cells.The relevant results are as following:1.The lentiviral plasmid plvx-zsgreen-sh MSL1 has been constructed successfully.The best infection titer is 0.6×virus and the best infection time is 72 h.We use Western Blot and RT-PCR to detect the plasmid can interfere with the expression of MSL1 at protein and m RNA level.2.CCK-8 and colony formation experiments prove that knocking down the MSL1 gene can inhibit the survival and proliferation of SW480 and SW620 colon cancer cells.3.Hoechst staining experiment and Western Blot method prove that knocking down the MSL1 gene can promote the apoptosis of SW480 and SW620 colon cancer cells.4.The low expression of MSL1 in cells can lead to low levels of E-cadherin and increase the expression of N-cadherin,suggesting that MSL1 may be involved in the EMT process.5.After knocking down MSL1,the phosphorylation levels of ERK,Akt and mTOR proteins in the MEK/ERK and PI3K/Akt/mTOR signal pathways are significantly reduced,suggesting that MSL1 may effect the process pf proliferation and apoptosis of colcon cancers by regulating the above signal pathways.This experiment provides reference value for studying the occurrence and development of colon cancer at the level of epigenetics,and it is conducive to followup treatment,mainly because an original direction for the treatment of colon cancer patients can be provided.