摘要
目的探究环状RNA hsa_circ_0006135在胃癌增殖、侵袭以及细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)中的作用,及其影响胃癌进展的具体机制。方法收集江苏大学附属人民医院普外科收治的45名胃癌患者的癌组织及癌旁正常组织标本,实时荧光定量PCR(qRT-PCR)检测hsa_circ_0006135在胃癌组织及胃癌细胞株(HGC-27和MGC-803)中的表达情况。体外培养胃癌细胞株HGC-27和MGC-803,分别敲减和(或)过表达hsa_circ_0006135、miR-1236-3p,克隆形成实验检测细胞增殖能力,Transwell小室检测细胞侵袭能力,并以Western blot检测各组细胞E-cadherin、Vimentin、N-cadherin、MTA2(metastasis-associated tumor gene family 2)蛋白表达情况。建立小鼠异种移植皮下瘤模型,观察敲减hsa_circ_0006135对胃癌细胞HGC-27体内成瘤的影响。结果hsa_circ_0006135在胃癌组织和胃癌细胞株HGC-27、MGC-803中高表达;敲减其表达后,胃癌细胞株的增殖、侵袭和EMT过程受到抑制。miR-1236-3p在HGC-27、MGC-803细胞株中低表达,hsa_circ_0006135可能作为miR-1236-3p的分子海绵发挥作用。MTA2在胃癌细胞株中高表达,miR-1236-3p可与MTA2相作用,并调节其表达。miR-1236-3p inhibitor可以逆转hsa_circ_0006135敲减对胃癌细胞增殖、侵袭、EMT过程的抑制作用,过表达MTA2则可以消除miR-1236-3p对胃癌细胞增殖、侵袭、EMT的负调控。敲减hsa_circ_0006135可在体内抑制移植瘤生长。结论hsa_circ_0006135在胃癌发展中起促进作用,可能通过调节miR-1236-3p/MTA2轴促进胃癌细胞的增殖、侵袭和EMT过程。
Objective The purpose of this study is to explore the role of hsa_circ_0006135in the proliferation,invasion and epithelial-mesenchymal transition(EMT)of gastric cancer,and the specific mechanism of its influence on the progression of gastric cancer.Methods The cancer tissue and normal tissue samples of 45gastric cancer patients admitted to the General Surgery Department of the People’s Hospital Affiliated to Jiangsu University were collected,and real-time fluorescent quantitative PCR(qRT-PCR)was used to detect the expression of hsa_circ_0006135in gastric cancer tissues and gastric cancer cell lines(HGC-27 and MGC-803).Gastric cancer cell lines HGC-27and MGC-803were cultured in vitro,and hsa_circ_0006135and miR-1236-3p were knocked down and/or overexpressed respectively.The cell proliferation ability was detected by colony formation assay,and the cell invasion ability was detected by Transwell chamber.The protein expressions of E-cadherin,Vimentin,N-cadherin and MTA2(metastasis-associated tumor gene family 2)were detected by Western blotting.A mouse xenograft subcutaneous tumor model was established to observe the in vivo tumorigenesis of gastric cancer cell line HGC-27with hsa_circ_0006135knocked down.Results hsa_circ_0006135was highly expressed in gastric cancer tissue as well as gastric cancer cell line HGC-27and MGC-803.After knocking down its expression,the proliferation,invasion and EMT process of gastric cancer cell lines were inhibited.miR-1236-3p was low expressed in HGC-27and MGC-803,and hsa_circ_0006135functioned as molecular sponge for miR-1236-3p.MTA2was highly expressed in gastric cancer cells,and miR-1236-3p had a targeting effect on MTA2.miR-1236-3p inhibitor reversed the inhibitory effect of hsa-circ_0006135silencing on gastric cancer cell proliferation,invasion,and EMT process.Overexpression of MTA2eliminated the negative regulation of miR-1236-3p on gastric cancer cell proliferation,invasion,and EMT.In vivo experiment showed that hsa_circ_0006135knockdown could inhibit growth of subcutaneous t
作者
高生宝
夏德鑫
张珍珍
田仪督
步雪峰
Gao Shengbao;Xia Dexin;Zhang Zhenzhen(School of Medicine,Jiangsu University,Zhenjiang 212013,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2023年第5期585-593,共9页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金面上项目(No.81672999)
镇江市重点研发计划项目(No.SH2023004)。
