摘要
目的探讨长链非编码RNA BC015134促进胃癌细胞侵袭迁移的作用及机制。方法通过对GEO数据库中10对胃癌和癌旁组织RNA芯片结果进行分析以及湖州市中心医院2015年1月至2020年12月胃癌根治术患者的临床样本80例验证,筛选胃癌转移相关长链非编码RNA。通过shRNA介导的RNA干扰和质粒介导的过表达技术调节BC015134在胃癌MGC-803细胞中的表达水平,采用Transwell实验检测胃癌细胞侵袭和迁移能力。通过RNA pulldown实验及Western blot法验证BC015134与β-连环蛋白(β-Catenin)能否相互结合。利用qRT-PCR技术检测BC015134在临床样本中的表达水平,并做预后分析。结果通过对芯片结果进行临床样本验证,发现BC015134在有远处转移的胃癌原发灶中高表达。在MGC-803细胞中过表达BC015134能够促进细胞的侵袭及迁移能力,而敲减BC015134能够抑制细胞的侵袭及迁移能力。BC015134能够与β-Catenin相互结合,过表达β-Catenin后神经型钙黏蛋白(N-cadherin)的表达上调,上皮型钙黏蛋白(E-cadherin)的表达下调。结论BC015134通过与β-Catenin相互结合,促进胃癌细胞上皮间质转化,从而促进胃癌细胞的侵袭与迁移。
Objective To investigate the effect and mechanisms of lncRNA BC015134 in the metastasis and invasion of gastric cancer cells.Methods Through the analysis of the results of 10 pairs of gastric cancer and para-cancer chips in the GEO database and the validation of 80 clinical samples collected in Huzhou Central Hospital from January 2015 to December 2020,the long chain non-coding RNAs(lncRNAs)related to gastric cancer metastasis was screened.The expression levels of BC015134 in gastric cancer cells was regulated by shRNA mediated overexpression and RNAi methods.The Transwell method was adopted to explore the migration and invasion functions of BC015134 in gastric cells.RNA pull-down assay combined with Western blot analysis were used to validate the combination of BC015134 withβ-Catenin.The expression levels of BC015134 in clinical samples were analyzed by qRT-PCR.Results LncRNA BC015134 was highly expressed in primary tumors with distant metastasis compared with those without metastasis.Overexpressing BC015134 enhanced the migration and invasion ability of gastric cancer MGC-803 cells while knocking down of BC015134 had the opposite effects.BC015134 was able to bind toβ-Catenin leading to the overexpression of N-cadherin and down expression of E-cadherin.Conclusion LncRNA BC015134 can induce epithelial-mesenchymal transition of gastric cancer cells by binding toβ-Catenin,to promote the migration and invasion of gastric cancer cells.
作者
邬仲鑫
蔡炜龙
尹磊
WU Zhongxin;CAI Weilong;YIN Lei(Department of Gastrointestinal Hernia Surgery,Huzhou Central Hospital(Affiliated Huzhou Hospital Zhejiang University School of Medicine),Huzhou 313000,China)
出处
《浙江医学》
CAS
2023年第3期235-238,250,I0004,共6页
Zhejiang Medical Journal
基金
浙江省医药卫生科技计划项目(2019ZD051)。
作者简介
通信作者:尹磊,E-mail:15056997829@163.com。
