MYH10 Induces MYH9 To Promote Ovarian Cancer Cell Proliferation,Migration And Invasion

Background and ObjectivesOvarian cancer is one of the most common malignant cancers in female reproductive system,and it is also the highest mortality rate of the female reproductive system.Although the prognosis of ovarian cancer patients has improved with the improvement of various clinical treatments(including surgery,chemotherapy,PARP inhibitors,anti-angiogenesis drugs,etc.),but the survival rate of advanced patients is still not optimistic.The pathogenesis of ovarian cancer is yet unclear.Its occurrence may be related with age,fertility,blood type,mental factors,hormones,genetics,the environment,and so on.The cause of death is mainly related with the high degree of malignancy and recurrence,metastasis of ovarian cancer.Therefore,it is of great value to explore the molecular mechanism and biological events of ovarian cancer,which will obviously improve the prognosis and overall survival rate of ovarian cancer patients.MYH10 is encoded by non-smooth myosin heavy chain ?B.It plays a different role in different cancers.It plays an oncogenic role in lung cancer,invasive bladder cancer,and myelogenous leukemia,which promotes cell proliferation,invasion and metastasis of these cancers.In contrast,MYH10 negatively regulates cell growth and proliferation in breast epithelial cells.However,there is few report on the expression,specific functions and molecular mechanisms of MYH10 in ovarian cancer.MYH9,also known as NMMHC?A,encodes a non-smooth myosin heavy chain.Recently,many studies have reported that it is involved in the pathogenesis of many cancers.It has been reported that it plays an oncogenic role in promoting tumor proliferation,invasion and metastasis in gastric cancer,colorectal cancer,breast cancer,acute myeloid leukemia,esophageal cancer,lung cancer,and so on.However,in contrast,MYH9 plays a tumor suppressor role in squamous cell carcinoma by regulating the stability of p53.However,there is few report about MYH9 expression,specific function and molecular mechanism in ovarian cancer.In this study,we analyzed the expression,role,and mechanism of MYH10 and MYH9 in ovarian cancer,respectively,and further discussed the coexpression and interaction between MYH10 and MYH9,to confirm that MYH10 can induce MYH9 expression to promote proliferation,invasive,and metastasis of ovarian cancer by regulating Wnt/?-catenin signaling pathways.This study will provide novel ideas for the progression of ovarian cancer.Contents and methods1.The expression,function and mechanism of MYH10 in ovarian cancer(1)Immunohistochemical experiments were performed to detect the expression of MYH10 in epithelial ovarian cancer paraffin tissues and paratumor tissues,and to analyze the relationship between MYH10 and clinical parameters,prognosis of ovarian cancer patients,respectively;(2)The effects of MYH10 on the proliferation of ovarian cancer cells were observed with MTT,Edu and colony formation to determine the function of MYH10 in ovarian cancer SKOV3 and OVCAR3 cells;(3)The effects of MYH10 on the migration and invasion of ovarian cancer cells were detected by Transwell,Boyden and scratch experiments to determine the function of MYH10 in SKOV3 and OVCAR3 cells of ovarian cancer;(4)Western blot was performd to detect the effect of MYH10 on the expression of Wnt/?-catenin pathway factors(including P-catenin,EMT,stemness factors,and so on)to explore the mechanism of MYH10 in ovarian cancer.2.The expression,function and mechanism of MYH9 in ovarian cancer(1)Immunohistochemical experiments were performed to detect MYH9 expression in epithelial ovarian cancer paraffin and paratumor tissues,and to analyze the relationship between MYH9 and clinical parameters,prognosis of ovarian cancer patients,respectively;(2)The effects of MYH9 on the proliferation of ovarian cancer cells were detected with MTT and Edu experiments to determine the function of MYH9 in ovarian cancer SKOV3 and OVCAR3 cells;(3)The effects of MYH9 on the migration and invasion of ovarian cancer cells were detected with Transwell,Boyden and scratch experiments to determine the function of MYH9 in ovarian cancer SKOV3 and OVCAR3 cells;(4)Western blot was performed to detect the effect of MYH9 on the expression of Wnt/?-catenin pathway factors(including P-catenin,EMT,sternness factors,and so on)to explore the mechanism of MYH9 in ovarian cancer.3.MYH10 induces MYH9 to promote ovarian cancer cell proliferation,migration and invasion(1)Immunohistochemical experiments were performed to detect coexpression of MYH10 and MYH9 in 219 cases of epithelial ovarian cancer paraffin and 41 cases of paratumor tissues,and to analyze the correlation of MYH10 and MYH9,and to analyze the reltionship between MYH10/MYH9 and clinical parameters,prognosis of epithelial ovarian cancer patients,respectively;(2)MYH10 interacts and promotes MYH9 expression1)Bioinformatics software was used to predict that MYH9 is one of the candidate interacting proteins of MYH10;2)Western Blot was used to detect the expression of MYH9 in MYH10 Knockdown or overexpressed SKOV3 cells;3)Endogenous CoIP experiments were used to confirm that MYH9 is a direct interacting protein of MYH10;4)MTT,Edu,Transwell and scratch experiments were used to determine whether the addition of siMYH9 to MYH10 overexpressed ovarian cancer cells could reverse the function of MYH10 on cell proliferation,migration and invasion;5)Western blot was used to detect whether the transfection of siMYH9 into MYH10 overexpressed ovarian cancer cells could reverse the expression of Wnt/?-catenin signaling pathway.Results1.Oncogene of MYH10 can promote proliferation,migration and invasion of ovarian cancer cells(1)Immunohistochemical experiments showed that MYH10 was significantly higher in epithelial ovarian cancer than in paratumor tissues,and it was an independent prognostic factor in epithelial ovarian cancer patients;(2)The results of MTT,Edu and colony formation experiments showed that MYH10 could promote the proliferation of ovarian cancer SKOV3 and OVCAR3 cells;(3)Transwell,Boyden and scratch experiments showed that MYH10 could promote the migration and invasion of ovarian cancer SKOV3 and OVCAR3 cells;(4)Mechanism experiments showed that MYH10 could promote cell proliferation,migration and invasion of ovarian cancer by regulating Wnt/?-catenin and downstream pathway;2.Oncogene MYH9 can promote ovarian cancer cell proliferation,migration and invasion(1)Immunohistochemical results showed that MYH9 was significantly higher in epithelial ovarian cancer than in paratumor tissues,and it was an independent prognostic factor in epithelial ovarian cancer patients;(2)MTT and Edu experiments results showed that MYH9 could promote ovarian cancer SKOV3 and OVCAR3 cell proliferation;(3)Transwell,Boyden and scratch experiments showed that MYH9 could promote the migration and invasion of ovarian cancer SKOV3 and OVCAR3 cells;(4)Mechanism experiments showed that MYH9 could promote cell proliferation,migration and invasion of ovarian cancer by regulating Wnt/?-catenin and downstream pathway;3.MYH10 induces MYH9 to promote ovarian cancer cell proliferation,migration and invasion1)There was a significantly positive correlation between MYH10 and MYH9 in epithelial ovarian cancer;and coexpression of MYH10+/MYH9+ was an indeed dependent prognostic factor;2)Endogenous CoIP experiments confirmed the interaction between MYH10 and MYH9;3)Western blot experiments confirmed that the expression of MYH9 was inhi-bited in MYH10 knockdown SKOV3 cells;and MYH9 was elevated in MYH10 overexpressed SKOV3 cells.4)siMYH9 was transfected into MYH10 overexpressed ovarian cancer cells.MTT and Edu experiments showed that siMYH9 could reverse the proliferation function of MYH10 in ovarian cancer;Moreover,Transwell and scratch experiments showed that siMYH9 could reverse the migration and invasion of MYH10 in ovarian cancer;5)Western blot results showed that siMYH9 could reverse the regulatory effect of MYH10 on Wnt/?-catenin signaling pathway.Conclusions1.MYH10 expression is elevated in epithelial ovarian cancer tissues,and it can promote the proliferation,migration and invasion of ovarian cancer cells by regulating the Wnt/?-catenin signaling pathway;2.MYH9 expression is elevated in epithelial ovarian cancer tissues,and it can promote the proliferation,migration and invasion of ovarian cancer cells by regulating the Wnt/?-catenin signaling pathway;3.Coexpression of MYH10+/MYH9+ is an indeed dependent prognostic factor,and MYH10 induces MYH9 to promote proliferation,migration and invasion of ovarian cancer cells by regulating Wnt/?-catenin signaling pathway.