|Table of Contents|

Effects of GAS6-AS1 regulating miR-370-3p/SPATA2 axis on proliferation,migration,invasion,apoptosis and EMT of ovarian cancer cells

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2024 03
Page:
424-431
Research Field:
Publishing date:

Info

Title:
Effects of GAS6-AS1 regulating miR-370-3p/SPATA2 axis on proliferation,migration,invasion,apoptosis and EMT of ovarian cancer cells
Author(s):
JIA Yijuan; WANG Zhongxian; WANG Donghua; GONG Shixiong
Department of Gynecology,Wuhan First Hospital,Hubei Wuhan 430022，China.
Keywords:
long non-coding RNA; GAS6 antisense RNA1; miR-370-3p; spermatogenesis-associated protein 2; ovarian cancer; epithelial mesenchymal transformation
PACS:
R737.31
DOI:
10.3969/j.issn.1672-4992.2024.03.006
Abstract:
Objective:To investigate the impacts of long non-coding RNA GAS6 antisense RNA1(lncRNA GAS6-AS1) on proliferation,migration,invasion,apoptosis and epithelial mesenchymal transformation(EMT) of ovarian cancer cells by regulating miR-370-3p/spermatogenesis-associated protein 2(SPATA2) axis.Methods:qRT-PCR and Western blot were used to detect the expression of GAS6-AS1,miR-370-3p,and SPATA2 proteins in adjacent cancer tissues,ovarian cancer tissues,human normal ovarian epithelial cells IOSE80,and ovarian cancer cell lines HO-8910,SKOV3,and A2780,respectively.SKOV3 cells were grouped into:Control group(NC group),si-NC group,si-GAS6-AS1 group,mimic NC group,miR-370-3p mimic group,si-GAS6-AS1+inhibitor NC group,and si-GAS6-AS1+miR-370-3p inhibitor group,qRT-PCR was applied to detect the expression of GAS6-AS1 and miR-370-3p in cells.CCK-8 method was applied to detect cell proliferation.Flow cytometry was applied to detect cell apoptosis.Scratch healing test was applied to detect cell migration.Transwell experiment was applied to detect cell invasion.Western blot was applied to detect the protein expression of SPATA2,CyclinD1,Bcl-2-associated X(Bax),E-cadherin,Vimentin,and N-cadherin.Double Luciferase reporter gene experiment was applied to detect the relationship between GAS6-AS1 and miR-370-3p,and between miR-370-3p and SPATA2.Results:In ovarian cancer tissues and cells,GAS6-AS1 and SPATA2 proteins were highly expressed,while miR-370-3p was lowly expressed,and GAS6-AS1 and SPATA2 protein expression levels were the highest and miR-370-3p expression level was the lowest in SKOV3 cells,therefore,SKOV3 cells were selected as the follow-up research object.Compared with the NC group and si-NC group,the GAS6-AS1,OD450 value(24 h,48 h,72 h),scratch healing rate,number of invasive cells,expression of SPATA2,CyclinD1,Vimentin,N-cadherin proteins in the si-GAS6-AS1 group decreased,the expression of miR-370-3p,cell apoptosis rate,and expression of Bax and E-cadherin proteins increased(P＜0.05).Compared with the NC group and mimic NC group,the OD450 value(24 h,48 h,72 h),scratch healing rate,number of invasive cells,expression of SPATA2,CyclinD1,Vimentin,N-cadherin proteins in the miR-370-3p mimic group decreased,the expression of miR-370-3p,cell apoptosis rate,and expression of Bax and E-cadherin proteins increased(P＜0.05).miR-370-3p inhibitor weakened the inhibitory effects of silencing GAS6-AS1 on SKOV3 cell proliferation,migration,invasion,EMT,and the promoting effect on cell apoptosis.There was a targeted regulatory relationship between GAS6-AS1 and miR-370-3p,and between miR-370-3p and SPATA2.Conclusion:Silencing GAS6-AS1 inhibits SPATA2 expression by up-regulating miR-370-3p,thereby inhibiting SKOV3 cell proliferation,migration,invasion,EMT,and promoting cell apoptosis.

References:

［1］SUNG H,FERLAY J,SIEGEL RL,et al.Global cancer statistics 2020:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］.CA Cancer J Clin,2021,71(3):209-249.
［2］NING X,SHI G,REN S,et al.GBAS regulates the proliferation and metastasis of ovarian cancer cells by combining with eEF1A1［J］.Oncologist,2022,27(1):64-75.
［3］WU XP,XU ZQ,XIE WM,et al.Long non-coding RNA GAS6-AS1 enhances breast cancer cell aggressiveness by functioning as a competing endogenous RNA of microRNA-215-5p to enhance SOX9 expression［J］.Exp Ther Med,2022,23(1):109-115.
［4］WANG Y,DING M,YUAN X,et al.lncRNA SNHG15 promotes ovarian cancer progression through regulated CDK6 via sponging miR-370-3p［J］.Biomed Res Int,2021,2021(1):9394563-9394571.
［5］WIESER V,TSIBULAK I,DEGASPER C,et al.Tumor necrosis factor receptor modulator spermatogenesis-associated protein 2 is a novel predictor of outcome in ovarian cancer［J］.Cancer Sci,2019,110(3):1117-1126.
［6］GENG L,WANG Z,TIAN Y.Down-regulation of ZNF252P-AS1 alleviates ovarian cancer progression by binding miR-324-3p to downregulate LY6K［J］.J Ovarian Res,2022,15(1):1-15.
［7］NAROD S.Can advanced-stage ovarian cancer be cured［J］.Nat Rev Clin Oncol,2016,13(1):255-261.
［8］KIM LK,PARK SA,YANG Y,et al.LncRNA SRA mediates cell migration,invasion,and progression of ovarian cancer via NOTCH signaling and epithelial-mesenchymal transition［J］.Biosci Rep,2021,41(9):BSR20210565.
［9］ZHANG H,FENG X,ZHANG M,et al.Long non-coding RNA CASC2 upregulates PTEN to suppress pancreatic carcinoma cell metastasis by downregulating miR-21［J］.Cancer Cell Int,2019,19(1):18-25.
［10］CHEN Q,ZHOU L,MA D,et al.LncRNA GAS6-AS1 facilitates tumorigenesis and metastasis of colorectal cancer by regulating TRIM14 through miR-370-3p/miR-1296-5p and FUS［J］.J Transl Med,2022,20(1):356-370.
［11］徐华,张海萍,王虹伊,等.LncRNA GAS6-AS1靶向miR-324-3p激活Wnt/β-catenin通路调控脑胶质瘤细胞增殖及放疗敏感性的研究［J］.河北医药,2022,44(19):2885-2890. XU H,ZHANG HP,WANG HY,et al.Effects of lncRNA GAS6-AS1 on the proliferation and radiotherapy sensitivity of glioma T98G cells by targeting miR-324-3p to activate Wnt/β-catenin pathway［J］.Hebei Medical Journal,2022,44(19):2885-2890.
［12］ZHANG C,LI Z,WANG J,et al.Ethanol extracts of solanum lyratum thunb regulate ovarian cancer cell proliferation,apoptosis,and epithelial-to-mesenchymal transition(EMT) via the ROS-mediated p53 pathway［J］.J Immunol Res,2021,2021(1):5569354-5569369.
［13］CUI P,LI H,WANG C,et al.UBE2T regulates epithelial-mesenchymal transition through the PI3K-AKT pathway and plays a carcinogenic role in ovarian cancer［J］.J Ovarian Res,2022,15(1):103-115.
［14］ZHAO Q,ZHANG B,LI Z,et al.Effects of lncRNA PROX1-AS1 on proliferation,migration,invasion and apoptosis of lung cancer cells by regulating miR-1305［J］.J Healthc Eng,2022,2022(1):9570900-9570908.
［15］CHOI HS,LIM ES,BAEK KH.Deubiquitinating enzyme USP12 regulates the pro-apoptosis protein Bax［J］.Int J Mol Sci,2022,23(21):13107-13122.
［16］LI M,XIE Z,WANG P,et al.The long noncoding RNA GAS5 negatively regulates the adipogenic differentiation of MSCs by modulating the miR-18a/CTGF axis as a ceRNA［J］.Cell Death Dis,2018,9(5):554-566.
［17］赵志国，张力平.miRNA和lncRNA相互作用与恶性肿瘤相关性的研究进展［J］.现代肿瘤医学，2018,26（18）：2991-2994. ZHAO ZG,ZHANG LP.Advancements of interaction between miRNA and lncRNA on malignant tumor［J］.Modern Oncology,2018,26（18）：2991-2994.
［18］武红,李枫,张曦辉,等.miR-370-3p靶向HDAC4调节卵巢癌SKOV3细胞的生长和代谢研究［J］.中国肿瘤临床,2020,47(23):1194-1199. WU H,LI F,ZHANG XH,et al.MicroRNA-370-3p targets histone deacetylase 4 to regulate growth and metabolism of ovarian cancer SKOV3 cells［J］.Chinese Journal of Clinical Oncology,2020,47(23):1194-1199.
［19］CHEN TY,XIONG F,ZHANG Z,et al.Effects of LncRNA IGF2-AS gene on proliferation and apoptosis of lung cancer cells by targeting miR-370-3p［J］.Chinese Journal of Gerontology,2021,41(23):5308-5313.
［20］JI H,ZHANG L,ZOU M,et al.SPATA2 suppresses epithelial-mesenchymal transition to inhibit metastasis and radiotherapy sensitivity in non-small cell lung cancer via impairing DVL1/β-catenin signaling［J］.Thorac Cancer,2023,14(11):969-982.
［21］W VERENA,AA SAMIRA,S SUSANNE,et al.TNFα signalling predicts poor prognosis of patients with endometrial cancer［J］.Carcinogenesis,2020,41(8):1065-1073. （编校:张西敏）

Memo

Memo:
湖北省科学技术厅2021年省科技计划项目(编号：2021CFB576)
Last Update: 2023-12-29