The Role And Mechanism Of Metastasis-associated Protein 2in The Proliferation And Metastasis Of Cervical Cancer Cells

Cervical cancer is a common malignant tumor in women,and its incidence rate ranks second in the world.The incidence of cervical cancer in China is on the rise,and it has ranked the first among female genital malignant tumors in China.At present,the treatment of cervical cancer mainly includes surgical treatment and concurrent chemoradiotherapy,but the mortality and prognosis of cervical cancer patients have not been significantly improved.Therefore,in-depth study of the pathogenesis of cervical cancer is of great significance for the creation of more effective and precise treatment.In recent years,epigenetic changes have received extensive attention in cervical tumors.Lysine acetylation modification is an important means of epigenetic changes and is closely related to tumor diseases.Metastasis-associated protein 2(MTA2)can be used as an oncogene to promote proliferation,migration and invasion of various cancer cells.In addition,lysine acetylation modification is involved in the proliferative effect of MTA2 on colorectal cancer cells,so acetylation modification is closely related to MTA2 biological function.However,the specific role of MTA2 in cervical cancer is unclear.This study focused on the role and mechanism of MTA2 and its acetylation modification in the proliferation and invasion of cervical cancer cells,and obtained the following results:1.The level of MTA2 protein acetylation in cervical cancer tissues is significantly up-regulated.The whole proteomics and acetylation proteomic analysis of cervical cancer tissues and adjacent tissues by LC-MS/MS showed that the level of MTA2 protein in cervical cancer samples was significantly increased,and the acetylation level of lysine site 592 was significantly increased.Further Western blot and immunoprecipitation showed that the levels of MTA2 protein and acetylation in clinical cervical cancer tissues were significantly higher than those in adjacent tissues.2.MTA2 promotes proliferation of cervical cancer cells through acetylation of K592 locus.MTA2 K592 site mutation can inhibit the protein expression and acetylation level of cervical cancer cell line MTA2.The results of CCK-8 and Flow cytometry showed that MTA2 K592 mutation could inhibit the proliferation and cell cycle progress of HeLa cells promoted by MTA2,but could not affect C-33 A cells.Western blot analysis showed that MTA2 K592 mutation could inhibit the regulation of MTA2 on the cell cycle-related proteins of HeLa and C-33 A cells.In vivo experiments in nude mice showed that MTA2 treatment significantly up-regulated the expression and acetylation of MTA2 protein in HeLa cell xenografts and C-33 A cell xenografts,distinctly promoted the growth of HeLa cell xenografts and C-33 A cell xenografts,respectively.Compared with MTA2 treatment,MTA2 K592 R treatment down-regulated MTA2 acetylation level and protein level,also significantly inhibited the growth of human cervical carcinoma xenografts in nude mice.3.MTA2 inhibits cervical cancer cell apoptosis and promotes migration and invasion of cervical cancer cells through acetylation at K592 site.Hoechst staining,flow cytometry,Western blot,scratch assay and Transwell assay were used to detect apoptosis,migration,invasion and autophagy-related protein expression of cervical cancer cells.The results showed that MTA2 significantly decreased the apoptosis and autophagy of HeLa and C-33 A cells,inhibited the expression of pro-apoptosis-related proteins,significantly promoted cell migration and invasion;K592 mutation can counteract the inhibition of MTA2 on cell apoptosis and autophagy,moreover reduce the promotion of MTA2 on cell migration and invasion.4.Acetyltransferases CBP and Tip60 are involved in the acetylation of MTA2 at the K592 site.The results of co-immunoprecipitation showed that K592 site mutation reduced the acetylation of MTA2 by CBP and Tip60,and inhibited the protein expression of MTA2,which was consistent with the trend of CBP and Tip60 inhibitors.In summary,protein and acetylation levels of MTA2 in cervical cancer tissues were significantly higher than those in adjacent tissues.MTA2 promoted the proliferation and metastasis of cervical cancer cells through K592 acetylation.The results of this study provide experimental basis for further exploring the molecular regulatory mechanism of cervical cancer occurrence and development,and also provide a new potential target for the treatment of cervical cancer.