| Background:Epithelial ovarian cancer(EOC)is the disease with the highest mortality rate in female reproductive system malignancies.Due to the atypical early symptoms of ovarian cancer and the lack of effective screening methods in the early stage,most ovarian cancers are already in the advanced stage when they are diagnosed.Different pathological types of ovarian cancer are slightly different in terms of pathogenesis,biological behavior,histological morphology,clinical manifestations,treatment methods and prognosis.Epithelial ovarian cancer is the most common,accounting for more than 90%.At present,the mechanism of occurrence and development of ovarian epithelial tumors is still unclear.Long non-coding RNAs are a class of eukaryotic non-coding RNA molecules longer than 200 nt.LncRNA Abnormal expression of lnc RNA has been found in a series of diseases.Detection of lnc RNA expression in different cells or disease states can help to understand its function,reveal its mechanism of action,or identify effective molecular markers.Methods:First,the human ovarian surface epithelial cell line OSE cells and epithelial ovarian cancer cells(ES-2,OVCAR3,CAOV-3 and SK-OV-3 cells)were cultured and collected,and the expression of LncRNA ADAMTS9-AS1 was detected by q RT-PCR.Then,OVCAR3 and CAOV-3 cells were transfected with si RNA targeting LncRNA ADAMTS9-AS1 for 24 h to silence the expression of LncRNA ADAMTS9-AS1.Cells were then re-treated with 30 n M of the ferroptosis inhibitor Fer-1.Cellular Fe2+,iron and ROS levels were detected by iron level detection kit and ROS detection kit.The expression of GPX4 protein was detected by Western blotting.CCK-8 proliferation assay,clone formation assay and Transwell assay were used to detect the changes of cell proliferation ability,clone formation ability and migration ability.Next,we used lnc Locator to predict the subcellular localization of LncRNA ADAMTS9-AS1.Its downstream micro RNAs were predicted online using mi Randa-3.3a software.In addition,we performed a dual-luciferase assay to confirm the binding site between LncRNA ADAMTS9-AS1 and mi R-587.Next,we further investigated the effect of mi R-587 on ferroptosis and cell function in epithelial ovarian cancer.The effect of LncRNA ADAMTS9-AS1/micro RNA-587/SLC7A11 signaling pathway on the function of epithelial ovarian cancer cells was detected by CCK-8 proliferation assay,clone formation assay and Transwell assay.Results:Quantitative real-time polymerase chain reaction(q RT-PCR)and Western blotting results showed that the expression of LncRNA ADAMTS9-AS1 was increased in epithelial ovarian cancer cells;after si RNA transfection of LncRNA ADAMTS9-AS1,the expression of micro RNA-587 was up-regulated and the expression of SLC7A11 was down-regulated;however,,micro RNA-587 inhibitor reversed the expression of SLC7A11 in LncRNA ADAMTS9-AS1 knockout cells.Ferroptosis-related marker detection and cell function assays confirmed that knockdown of LncRNA ADAMTS9-AS1 inhibited epithelial ovarian cancer cell proliferation and migration by promoting ferroptosis.Overexpression of mico RNA-587 also promoted ferroptosis while inhibiting cell proliferation and migration in epithelial ovarian cancer cells.Furthermore,mico RNA-587 inhibitor reversed the effects of LncRNA ADAMTS9-AS1 silencing on ferroptosis and cellular function.In addition,dual-luciferase gene reporter assay and RNA immunopre-cipitation assay confirmed the role of LncRNA ADAMTS9-AS1/ micro RNA-587/SLC7A11 signaling pathway in ovarian cancer.Conclusions:In conclusion,our study found that the LncRNA ADAMTS9-AS1 inhibits ferroptosis by targeting the mi R-587/SLC7A11 signaling pathway in epithelial ovarian cancer,thereby promoting proliferation.Our study provides a new treatment idea for epithelial ovarian cancer. |
