| ObjectiveOvarian cancer patients almost can be diagnosed in the late,and are prone to drug resistance during chemotherapy,leading to no significant improvement in overall survival rate after treatment.So seeking new treatment has become a hot research direction to solve this thorngoing problem.At present,ferroptosis has gradually been confirmed in a variety of tumor progression with its inhibitory effect,the researchers further explored the potential therapeutic effect,ferroptosis inducer Erastin can increase the sensitivity of the ovarian cancer cells to chemotherapy drug resistance,together to improve the fatality rate of chemotherapy drugs,the bottleneck of the findings in the treatment of ovarian cancer era provides a new train of thought.Based on the current research results,the specific mechanism of the effect of Erastin on the proliferation of ovarian cancer cells is not clear yet.Therefore,this paper aims to explore the effect of Erastin on the proliferation of human ovarian cancer cells and its mechanism.Methods1、Cell counting kit-8(CCK-8)assay and colony formation assay were used to detect the effect of Erastin in different doses on human ovarian cancer cell lines SKOV3 and A2780 cells activity and proliferation.2、ROS and MDA assays were used to detect the effects of 10μM Erastin on the production of ROS and MDA of peroxidation in SKOV3 and A2780 cells.3、RT-q PCR and Western blot assays were used to detect the effect of10μM Erastin on GSK-3β m RNA and protein expression in SKOV3 and A2780 cells.4、After the expression of GSK-3β was silenced,the effects of 10μM Erastin on proliferation and peroxidation of SKOV3 and A2780 cells were detected by CCK-8 assay,colony formation assay and ROS assay.5、After GSK-3β expression was silenced,the effects of 10μM Erastin on NRF2 and HO-1 protein expression in SKOV3 cell were detected by Western blot.Results1、CCK-8 assay and colony formation assay showed that the proliferation activity of SKOV3 and A2780 cells in the different doses of Erastin were used to treat the experimental group was significantly lower than that in the control group(P<0.001).2、ROS and MDA assays showed that the ROS and MDA production of SKOV3 and A2780 cells in the 10μM Erastin treatment group was significantly higher than that in the control group(P<0.001).3、RT-q PCR and Western blot showed that the expression level of GSK-3βin SKOV3 and A2780 cells in the 10μM Erastin treatment group was significantly higher than that in control group(P<0.001).4、After GSK-3β silencing,Western blot confirmed that the level of GSK-3βprotein in siGSK-3β+Erastin group was lower than that in si NC+Erastin group.5、CCK-8,colony formation assay and ROS assay showed that compared with si NC+Erastin group,the cell survival rate and number of clone formation were increased in siGSK-3β+Erastin group,while the ROS production ratio was decreased(P<0.05).6、Western blot showed that the expressions of NRF2 and HO-1 were decreased in si NC+Erastin group compared with the Control group.Compared with the si NC+Erastin group,the expression of NRF2 and HO-1 were increased in the siGSK-3β+Erastin group.ConclusionsExperiments confirmed that,ferroptosis inducer Erastin could inhibit the cell activity and proliferative ability of human ovarian cancer cells SKOV3 and A2780.By inducing cell peroxidation,increasing the production of cell peroxidation products ROS and MDA,and inhibiting the proliferative activity of cancer cells.Further investigation of the mechanism showed that Erastin inhibited the antioxidant response of cancer cells by increasing the expression of GSK-3β,and reducing the expression of NRF2 and HO-1,which made the metabolic environment abnormal,inhibited the protective NRF2 /HO-1antioxidant pathway,and thus inhibited the antioxidant response of cancer cells.This effect was inhibited by knockdown of GSK-3β expression.These results suggest that Erastin can inhibit the proliferation of ovarian cancer cells through the GSK-3β/NRF2 pathway,regulating cellular oxidative metabolism. |
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