|Table of Contents|

Gabra3 promotes the invasion and migration of bladder cancer T24 cells through AKT/mTOR pathway

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

Issue:
2024 07
Page:
1208-1214
Research Field:
Publishing date:

Info

Title:
Gabra3 promotes the invasion and migration of bladder cancer T24 cells through AKT/mTOR pathway
Author(s):
LI Mingming; HAN Lizhong; WANG Yanan; LU Guanjun; LYU Zhiyong
Department of Urology,General Hospital of Ningxia Medical University,Ningxia Yinchuan 750000,China.
Keywords:
γ-aminobutyric acid receptor subunit alpha-3; bladder cancer; T24 cells; apoptosis; migration; invasion; AKT/mTOR signaling pathway
PACS:
R737.14
DOI:
10.3969/j.issn.1672-4992.2024.07.005
Abstract:
Objective:To explore the role of Gamma-aminobutyric acid receptor subunit alpha-3(Gabra3) gene on apoptosis,migration and invasion of bladder cancer T24 cells and its mechanism.Methods:The TCGA database was used to analyze the expression of Gabra3 gene in bladder cancer and the correlation between metastasis and survival.T24 cell lines with Gabra3 overexpression and Gabra3 mutation were established.According to the transfection plasmid,T24 cells were divided into blank T24 cells(Control),T24 cells transfected with empty pDONR223 vector(NC),and T24 cells with wild type Gabra3 overexpression(wt-Gabra3),mutant Gabra3 T24 cell line(mut-Gabra3).In the complementation experiment,T24 cells were divided into mutant Gabra3(mut-Gabra3),mut-Gabra3 transfected with empty pDONR223 vector mut-Gabra3 group(mut-GABRA3-NC),and wild type Gabra3 overexpression group(mut-Gabra3+wt-Gabra3).The apoptosis of T24 cells was detected by TUNEL staining,the migration and invasion of T24 cells were detected by cell scratch and Transwell,respectively.The expression levels of AKT/mTOR pathway related molecules were detected by Western blot.Results:Gabra3 gene was significantly overexpressed in bladder cancer(P＜0.05),and the survival curve confirmed that patients with distant metastasis had a significantly shorter survival(P＜0.05).T24 cell lines with Gabra3 overexpression and mutation were successfully constructed.Compared with the Control group,the apoptosis ability of wt-Gabra3 group was significantly decreased,and the migration and invasion ability were significantly increased(P＜0.05),while the mut-Gabra3 group showed significantly increased apoptosis and decreased invasion(P＜0.05).The expression levels of p-AKT,p-mTOR,p-4E-BP1 and p-S6K proteins in wt-Gabra3 group were increased(P＜0.05),while mut-Gabra3 group had no difference in the above proteins.In complementation assay,the mut-Gabra3 mutant T24 cells overexpressing wt-Gabra3 showed inhibited apoptosis(P＜0.05),and the migration and invasion abilities were significantly enhanced(P＜0.05).AKT/mTOR pathway related molecules were significantly activated(P＜0.05).Conclusion:Exogenous unedited Gabra3 can promote the migration and invasion of bladder cancer T24 cells,which may be closely related to the activation of AKT/mTOR pathway.

References:

［1］ 温波,刘奔,夏露.非肌层浸润性膀胱癌的外科治疗选择［J］.临床泌尿外科杂志,2020,35(9):748-751. WEN B,LIU B,XIA L.Surgical treatment options for non-muscle invasive bladder cancer［J］.J Clin Urology,2020,35(9):748-751.
［2］ BHATTACHARAY D,GAWALI VS,KALLAY L,et al.Therapeutically leveraging GABA(A) receptors in cancer［J］.Exp Biol Med(Maywood),2021,246(19):2128-2135.
［3］ YANG Y,REN L,LI W,et al.GABAergic signaling as a potential therapeutic target in cancers［J］.Biomed Pharmacother,2023,161:114410.
［4］ ZHANG X,ZHANG R,ZHENG Y,et al.Expression of gamma-aminobutyric acid receptors on neoplastic growth and prediction of prognosis in non-small cell lung cancer［J］.J Transl Med,2013,11:102.
［5］ 田男.Gabra-3基因A至ⅠRNA编辑位点的鉴定及其分子机制的研究［D］.杭州：浙江大学,2010. TIAN N.Identification and molecular mechanism of Gabra-3 RNA editing sites from A to Ⅰ［D］.Hangzhou:Zhejiang University,2010.
［6］ DANIEL C,VENO MT,EKDAHL Y,et al.A distant cis acting intronic element induces site-selective RNA editing［J］.Nucleic Acids Res,2012,40(19):9876-9886.
［7］ GUMIREDDY K,LI A,KOSSENKOV AV,et al.The mRNA-edited form of GABRA3 suppresses GABRA3-mediated Akt activation and breast cancer metastasis［J］.Nat Commun,2016,7:10715.
［8］ PATIL V,PAL J,MAHALINGAM K,et al.Global RNA editome landscape discovers reduced RNA editing in glioma:Loss of editing of gamma-amino butyric acid receptor alpha subunit 3(GABRA3) favors glioma migration and invasion［J］.PeerJ,2020,8:e9755.
［9］SILVESTRIS DA,PICARDI E,CESARINI V,et al.Dynamic inosinome profiles reveal novel patient stratification and gender-specific differences in glioblastoma［J］.Genome Biol，2019,20(1):33.
［10］ 施言,王少慧,张发明.γ-氨基丁酸及其受体在肿瘤发生发展中的作用［J］.世界华人消化杂志,2012,20(5):45-50. SHI Y,WANG SH,ZHANG FM.Role of γ-aminobutyric acid and its receptors in tumorigenesis and development［J］.World Chinese Journal of Digestion,2012,20(5):45-50.
［11］ LIU Y,LI YH,GUO FJ,et al.Gamma-aminobutyric acid promotes human hepatocellular carcinoma growth through overexpressed gamma-aminobutyric acid A receptor alpha 3 subunit［J］.World J Gastroenterol，2008,14(47):7175-7182.
［12］LI YY,ZHENG XH,DENG AP,et al.MiR-92b inhibited cells EMT by targeting Gabra3 and predicted prognosis of triple negative breast cancer patients［J］.Eur Rev Med Pharmacol Sci，2019,23(23):10433-10442.
［13］ LONG M,ZHAN M,XU S,et al.MiR-92b-3p acts as a tumor suppressor by targeting Gabra3 in pancreatic cancer［J］.Mol Cancer，2017,16(1):167.
［14］ LIU J,WANG F,ZHANG Y,et al.ADAR1-mediated RNA editing and its role in cancer［J］.Front Cell Dev Biol,2022,10:956649.
［15］ JAIN M,JANTSCH MF,LICHT K.The editor's I on disease development［J］.Trends Genet,2019,35(12):903-913.
［16］ OHLSON J,PEDERSEN JS,HAUSSLER D,et al.Editing modifies the GABA(A) receptor subunit alpha3［J］.RNA,2007,13(5):698-703.
［17］ YU L,WEI J,LIU P.Attacking the PI3K/Akt/mTOR signaling pathway for targeted therapeutic treatment in human cancer［J］.Semin Cancer Biol,2022,85:69-94.
［18］陈康,邢基,张云龙,等.PI3K/AKT/mTOR通路抑制剂在膀胱癌中的研究进展［J］.现代肿瘤医学，2023,31（3）：575-579. CHEN K,XING J,ZHANG YL,et al.Research progress of PI3K/AKT/mTOR pathway inhibitors in bladder cancer［J］.Modern Oncology，2023,31（3）：575-579.
［19］曹李,陈鹏,李丁昌，等.p-mTOR、p-4EBP1及p-S6K1在结直肠腺癌组织中的表达及临床意义［J］.中国临床解剖学杂志,2022,40(5):541-547. CAO L,CHEN P,LI DC,et al.Expression and clinical significance of p-mTOR,p-4EBP1 and p-S6K1 in colorectal adenocarcinoma［J］.Chin J Clinical Anatomy,2022,40(5):541-547.
［20］ LIU L,YANG C,SHEN J,et al.GABRA3 promotes lymphatic metastasis in lung adenocarcinoma by mediating upregulation of matrix metalloproteinases［J］.Oncotarget,2016,7(22):32341-32350.

Memo

Memo:
宁夏自然科学基金（编号：2021AAC03325）
Last Update: 2024-02-29