Effects Of Harmine On Apoptosis And Invasion Of Ovarian Ancer Cells Through Wnt/β-catenin Signal Pathway

Research background and Objective:The ovaries are located deep in the pelvic cavity,and the incidence of ovarian cancer is hidden,there are no obvious symptoms in the early stage.Therefore,the early diagnosis of ovarian cancer(OC)is difficult,and more than 70% of epithelial ovarian cancer is diagnosed as advanced.The invasive ability of ovarian cancer is very strong.With the continuous progress of ovarian cancer,ovarian cancer can metastasize to some organs of pelvic cavity and abdominal cavity,and is prone to extensive metastasis in pelvic and abdominal cavity,including peritoneum,surface of abdominal visceral organs,greater omentum,diaphragm and retroperitoneal lymph nodes.Although about 80% of patients are sensitive to platinum chemotherapy drugs,but most patients gradually develop resistance with the increase of treatment,and the vast majority of patients are difficult to avoid recurrence and metastasis.Therefore,the in-depth study of the mechanism of occurrence,invasion,drug resistance and metastasis of ovarian cancer and the study of new anti ovarian cancer drugs are of great significance for the diagnosis and treatment of ovarian cancer.The antitumor efficacy of natural drugs and their extracts has become a research hotspot.The preliminary experiments of my graduate tutor team’s have confirmed that harmine can inhibit the proliferation and migration of ovarian cancer cells,and MTT tests have been conducted in the experiment that Harmine has a significant inhibitory effect on SKOV3 cells a t 10μ M and 24 hours,but the specific molecular mechanism is steel unknown.Studies have shown that many tumorigenesis,disease progression,poor prognosis and increased cancer-related mortality are closely related to the abnormal activation of Wnt/β-catenin signal,and the effective inhibition of Wnt/β-catenin signaling pathway may provide more options for the treatment of various types of cancer.This study was prepared from the tumor classical signaling pathway Wnt/β-catenin signaling pathway,focuses on the effect of harmine on the apoptosis and invasion of ovarian cancer,and deeply discusses its molecular mechanism.It is expected that this subject can further elaborate the specific mechanism of antitumor effect harmine,and provide new ideas for the diagnosis and treatment of ovarian cancer and drug research and development.Methods:1.Ovarian cancer SKOV3 cells were cultured in vitro and divided into two groups: dechromine group and control group.The dechromine group was treated with 10μM dechromine for 24 h,while the control group was treated with DMSO without dechromine(0μM).2.After co culture for 24 hours,the cells were stained with FITC-AV/PI,and the effect of harmine on apoptosis of ovarian cancer cells was analyzed by flow cytometry.3.Transwell chamber experiment was used to study the effect of harmine on the invasion of ovarian cancer cells.4.Western blot and qRT-PCR were used to detect the effect of harmine on proteins and genes related by Wnt/ β-catenin signaling pathway and apoptosis.5.In order to further verify whether harmine acts on Wnt/β-catenin signaling pathway to inhibit the invasion of ovarian cancer,and whether can reverse the effect of harmine on the invasion of ovarian cancer cells by activate the Wnt/β-catenin signaling pathway.We divided the cells into Control group(untreated,with the same concentration of DMSO only),Harmine group,and Harmine+lithium chloride(Li Cl)group.SKOV3 cells in the Control group were not treated,and SKOV3 cells in the Harmine group were treated with 10μ M Harmine for 24,SKOV3 cells in Harmine+Li CL group were treated with10μM Harmine and 10 mm Wnt/β-catenin signal pathway activator Li CL for 24 h.After 24 hours,the cell invasion function of each group was detected by Transwell chamber experiment.6.The measurement data were expressed in X±s,and were analyzed by prism 8software.The difference was statistically significant when p < 0.05.Results:1.The number of apoptosis of ovarian cancer cells in the harmine group was significantly higher than that in the control group.It is suggested that harmine can promote the apoptosis of ovarian cancer cells.2.The invasive ability of ovarian cancer cells in the harmine group was significantly weaker than that in the control group.It is suggested that harmine can inhibit the invasion of ovarian cancer cells.3.The expression of apoptosis related proteins Bax and cytochrome C in harmine group was stronger than that in control group,but the expression of Bcl-2protein was weaker than that in control group.The expression of Wnt/β-catenin signaling pathway related protein β-catenin,Cyclin D1 and c-myc protein in harmine group were weaker than that in the control group.The corresponding gene expression levels of the three proteins in the two groups were consistent with the protein expression results.4.After been activation of Wnt/β-catenin signaling pathway by lithium chloride,the cell invasion ability in harmine + Li Cl group was significantly higher than that of harmine group.Conclusion:1.Harmine can promote the apoptosis of ovarian cancer cells.2.Harmine can inhibit the invasion of ovarian cancer cells.3.Harmine may inhibits the invasion of ovarian cancer cells by inhibit Wnt/β-catenin signaling pathway.