The Influence Of MAP2K4 In Endometrial Cancer On Proliferation,invasion And Metastasis

Background and ObjectivesEndometrial cancer is an epithelial malignant tumor that origins from the endometrium and occurs in perimenopausal and postmenopausal women.Endometrial cancer is one of the most common female reproductive system tumors,and is the third leading cause of death of common gynecological malignancies just after ovarian cancer and cervical cancer.The etiology of endometrial cancer is complex.According to the literature,most of the endometrial cancer is due to endocrine disorders,and long-term sustained estrogen stimulation is an important factor in the pathogenesis of endometrial cancer.And the genetic factor and environmental are also have certain relationships.Endometrial cancer patients are often associated with obesity,infertility,menstrual abnormalities,polycystic ovary syndrome,postmenopausal delay,hypertension,diabetes and other factors.Mitogen-activated protein kinase kinase 4(MAP2K4)is a member of the MAP kinase signalling family.It has been reported that human mitogen-activated protein kinase kinase 4(MAP2K4)is encoded by human chromosome 17p11.2 and is one of the gene-targeted candidate tumor suppressor genes.The expression deletion of MAP2K4 gene has been found in the pancreas cancer,prostate cancer and ovarian cancer.And the down-regulation of the MAP2K4 gene may leading it development into a more aggressive phenotype that tends to invade and metastasize and become more resistant to chemistry Sexual irritation and difficulty in inactivation.In this study,we demonstrated that MAP2K4 inhibits the proliferation,invasion and migration of endometrial carcinoma,and which is produced by the PI3K/AKT-TGF-? signaling pathway.Together these results provide a molecular mechanism for the development and progression of endometrial carcinoma.Contents and methods1.To establish overexpression of MAP2K4 gene endometrial cancer cell lines:Ishikawa-MAP2K4 and RL95-2-MAP2K4.2.MTT,EDU,platelet cloning,cell cycle,subcutaneous tumor formation in nude mice were used to detect the proliferation of MAP2K4.3.Transwell,Boy den,scratches assays to detect the migration and invasion ability of MAP2K4.4.Western Blot was used to explore the molecular mechanism of MAP2K4 on endometrial carcinoma.I detected the cell cycle,EMT,PI3K/AKT-related signal pathway proteins to find the related molecular mechanism.Results1.MAP2K4 inhibit cell growth in EC.Using MTT assay,Edu incorporation assay,colony formation and cell cycle analysis,we found that overexpressed MAP2K4 significantly suppressed cell growth and G1 to S cell cycle transition in Ishikawa and RL95-2 cells.The mice injected with Ishikawa-MAP2K4 and RL95-2-MAP2K4 cells had smaller tumor burdens and displayed lower expression of Ki67 and PCNA in tumor tissues relative to controls.These results suggested MAP2K4 significantly inhibits tumorigenesis in vivo.The mechanism studies showed that Ishikawa-MAP2K4 and RL95-2-MAP2K4 decreased the expression of c-Myc,CDK4 and CCND1 and enhanced the expression of p27 and p15.In addition,we found that p-PI3K,p-AKT in Ishikawa-MAP2K4 and RL95-2-MAP2K4 expression decreased.2.MAP2K4 promote cell migration and invasion in EC.Transwell,scratches,Boyden assays indicated that MAP2K4 inhibits cell migration and invasion.Western blot showed that Ishikawa-MAP2K4 and RL95-2-MAP2K4 up-regulated E-cadherin,and down-regulated N-cadherin,TGF-?,vimentin,snail.We presumed that MAP2K4 may inhibit the EMT progression to inhibit the endometrial cancer.3.Clinical associations of MAP2K4 in EC.In 128 cases of endometrial carcinoma tissues,MAP2K4 was negatively correlated with FIGO stage and lymph node metastasis(p=0.010,p=0.016).Survival analysis showed that patients with high expression of MAP2K4 have a longer survival time.ConclusionAfter overexpress MAP2K4 gene into endometrial cancer cell Ishikawa and RL95-2,we conducted many in vitro and in vivo experiments which related to cell proliferation and invasion and metastasis,and we also did Western blot experiments.The results of all the above experiments showed that MAP2K4 inhibited the growth and invasion and metastasis of endometrial carcinoma cells.So we hypothesize that this process is mediated by PI3K/AKT and TGF-? signaling pathways.