The Role Of MYH9 In Promoting The Invasion And Metastasis Of Ovarian Cancer

BackgroundThe incidence of ovarian cancer ranks third among female reproductive system tumors.Its symptoms of early patients are not obvious.Ovarian cancer patients have a high recurrence rate and poor prognosis after treatment,and due to the diverse pathological types,the mechanism and etiology are still unclear.Myosin Heavy Chain 9(MYH9)is a widely expressed cytoplasmic myosin.Studies have found that MYH9 is highly expressed in many malignant solid tumors,and patients with high MYH9 expression have worse survival outcomes.MYH9 may play an oncogene role in tumor cell adhesion,migration,proliferation and differentiation,epithelial-mesenchymal transition(EMT)and other processes.Autophagy-dependent ferroptosis is a form of regulated cell death discovered in recent years,which has a great impact on cancer cells with a high tendency to metastasize.The NRF2 signaling pathway plays an important role in it.Promoting cancer cell death by regulating autophagy-dependent ferroptosis may be a new idea for suppressing tumor growth.Research purposesTo verify the role of MYH9 in the invasion and metastasis of ovarian cancer cells,to explore the relationship between MYH9 gene knockout and autophagy-dependent ferroptosis in ovarian cancer,and to further explore the signaling pathway to explore whether MYH9 may become a therapeutic agent for ovarian cancer target.Research method1.To explore the relationship between MYH9 and survival prognosis of ovarian cancer patients in public database.2.To compare expression of MYH9 in ovarian tissue samples by immunohistochemistry.3.To detect the expression difference of MYH9 in different ovarian cancer cells by RTqPCR,western blot and immunocytochemistry.4.To knock out ES-2 ovarian cancer cells with high MYH9 expression by lentiviral transfection,and verify its infection efficiency by immunofluorescence,RT-qPCR and Western blot.5.To detect the differences in cell proliferation ability,clone formation capabilities,apoptosis capabilities,cell migration and invasion capabilities in vitro,the tumorigenic ability,respectively,in the way of cell cycle,CCK8 assays,plate cloning assays,flow cytometry,transwell chamber assays and subcutaneous transplantation tumour model in nude mice.6.To detect the EMT-related indicators in cells by RT-qPCR and western blot,and the effect of MYH9 gene knockout on the EMT process was discussed.7.To observe the ultrastructure of cells by transmission electron microscopy,and preliminarily explore the effect of MYH9 gene knockout on autophagy-dependent ferroptosis and gene expression.8.To detect the effects of MYH9 gene knockout on the expression of ferroptosis and autophagy-related genes by RT-qPCR and Western blot,and explore the relationship with NRF2-related signaling pathways.Research result1.Public database shows that patients with high MYH9 expression have poor prognosis.2.At mRNA level and protein level,the expression of MYH9 in four different ovarian cancer cell lines is:ES-2>SKOV-3>CAOV-3>OVCAR-3.3.In the transfected ES-2 sgRNA cells,the expression of MYH9 was significantly reduced,the cell proliferation ability,clone formation ability,invasion and migration ability,tumorigenic ability in vitro were decreased,and the apoptosis ability increased.4.After MYH9 gene knockout,EMT-related indexes ZEB1,N-cadherin,β-catenin,Vimentin,Slug and Snail expression decreased in ovarian cancer cells,whereas E-cadherin expression increased.EMT process was inhibited.5.Electron microscope observation showed that the morphology of cells and mitochondria in ES-2 sgRNA group changed,autophagosome increased,and autophagy-dependent ferroptosis occurred at the same time.RT-qPCR and Western blot experiments also confirmed this result,that is,MYH9 gene knockout promoted autophagy-dependent ferroptosis.Conclusion and significance1.MYH9 is highly expressed in human ovarian cancer tissues and is associated with poor survival outcomes of patients.2.MYH9 gene knockout can inhibit the migration,invasion,proliferation,clone formation,tumorigenesis of ovarian cancer cells in vitro,and inhibit the process of EMT.3.MYH9 may target the NRF2 signaling pathway to inhibit autophagy-dependent ferroptosis.