Effects And Mechanism Of ?-tocopherol On The Proliferation And Migration Of SKVO3 Cells

Objective:The most common ovarian malignant tumor is ovarian epithelial cancer.Due to its hidden early symptoms and the relative lack of screening methods,most patients have advanced tumors when they are diagnosed.According to data from the National Cancer Center,the 5-year overall survival rate of ovarian cancer has hovered at 30%in the past 20 years.Tumor cytoreductive surgery supplemented with platinum-based combination chemotherapy is often used as the standard initial treatment for advanced ovarian cancer.Although pre-treatment can remit most patients,almost all patients with advanced ovarian cancer will face disease recurrence.In addition,the presence of gonadotropins and estrogen in ovarian cancer can increase the production of ROS,and then up-regulate the expression of Nrf2,which affects the biological behavior of tumor cells such as proliferation,apoptosis and migration.Tocopherol is a common non-enzymatic antioxidant that can effectively correct the imbalance of the redox system in tumor cells,and thus plays an important role in the occurrence and development of tumors.But in recent years,related reports of ?-tocopherol have different roles in different cancers,so it is worth exploring the biological role and possible mechanism of ?-tocopherol in ovarian cancer.This study aimed to explore the effects of ?-tocopherol treatment on the proliferation and migration of SKVO3 cells in ovarian cancer SKVO3 cell line;and to further explain its role and the expression of pan-PKC,ERK1/2,YAP1,Nrf2,Keap1 protein.Methods:1.Treat ovarian cancer SKVO3 cell line with ?-tocophero1 25mg/L,50mg/L,100mg/L,200mg/L and 400mg/L respectively,and set up DMSO treatment group as negative control group.CCK-8 solution was used to measure the cell proliferation of each group at 24h and 48h after treatment.2.Treat ovarian cancer SKVO3 cell line with ?-tocopherol 25mg/L,50mg/L,100mg/L and 200mg/L respectively,and set up DMSO treatment group as negative control group.At 0h,each group was subjected to scratch treatment and the width of the scratch was recorded,and the migration of SKVO3 cells at the scratch was measured at 24h and 48h after the scratch treatment.3.Treat ovarian cancer SKVO3 cell line with ?-tocopherol 25mg/L,50mg/L,100mg/L and 200mg/L respectively,and set up DMSO treatment group as negative control group.At 0h,SKVO3 cells of each group were plated on the Transwell respectively.After 24h,after staining with crystal violet,the number of SKVO3 cell migration in each group was observed.4.Using the genetic information in the Oncomine Database to clarify the expression of YAP 1 and Nrf2 in the tissues or blood of patients with ovarian cancer compared with normal human ovarian tissues or blood.Using big data analysis to screen ovarian cancer patient information in the TCGA database,YAP1 and Nrf2 were screened into high and low expression groups,and Kaplan-Meier was used to describe the relationship between YAP1 and Nrf2 expression levels and the survival prognosis of ovarian cancer patients.5.Treat ovarian cancer SKVO3 cell line with ?-tocopherol 25mg/L,50mg/L,100mg/L and 200mg/L respectively,and set up a DMSO treatment group as a negative control group.After 48h of treatment,the expression of pan-PKC,ERK1/2,YAP1,Nrf2,Keap1 protein in each group was measured by western blot.Results:1.CCK-8 experiment showed that:?-tocopherol at dose of 25 mg/L,50 mg/L,100 mg/L,200 mg/L,and 400 mg/L inhibited SKVO3 cells proliferation respectively,and so did differently at time of 24h,48h and 72h,P<0.05.2.The cell scratch test showed that ?-tocopherol had no effect on the migration of SKVO3 cells at whatever dose(25 mg/L,50 mg/L,100 mg/L,and 200 mg/L)and whatever time(24h and 48h),P>0.05.3.Transwell experiment showed that ?-tocopherol had no effect on the migration of SKVO3 cells at whatever dose(25 mg/L,50 mg/L,100 mg/L,and 200 mg/L)and after 24h,P>0.05.4.There was no different expression of YAP 1 between ovarian cancer tissue and normal ovarian tissue(P=0.290).The expression of Nrf2 in the blood of normal people was lower than ovarian cancer patients,P<0.05.5.SKVO3 cells were treated by ?-tocopherol at dose of 25mg/L,50mg/L,100mg/L and 200mg/L,respectively.48h later,the protein expression levels of pan-PKC,YAP1,Nrf2,ERK1/2 were decreased with ?-tocopherol dose increasing(P<0.05),on the contrary,the protein expression level of Keaplwas increased(P<0.05)Conclusions:1.?-tocopherol can inhibit the proliferation of ovarian epithelial SKVO3 cells in a time-and dose-dependent manner,but has no effect on its migration.2.?-tocopherol may decrease the expression of pan-PKC,YAP1,Nrf2,and ERK1/2 proteins,and increase the expression of Keap1;thereby affecting the proliferation of SKVO3 cells.