中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (7): 497-504.doi: 10.19401/j.cnki.1007-3639.2020.07.003

• 论著 • 上一篇    下一篇

STC1诱导上皮-间质转化促进肺癌细胞的侵袭和迁移

魏丽荣 1 ,滕小艳 1 ,夏前林 1 ,杜玉珍 1,2   

  1. 1. 上海健康医学院附属第六人民医院东院检验科,上海 201306 ;
    2. 上海市第六人民医院东院检验科,上海 201306
  • 出版日期:2020-07-30 发布日期:2020-08-06
  • 通信作者: 杜玉珍 E-mail: duyuzhen2005@163.com
  • 基金资助:
    上海健康医学院种子基金(HMSF-17-22-024)。

STC1 induces epithelial-mesenchymal transition to promote invasion and migration of lung cancer cells

WEI Lirong 1 , TENG Xiaoyan 1 , XIA Qianlin 1 , DU Yuzhen 1,2   

  1. 1. Department of Laboratory Medicine, Shanghai Sixth People’s Hospital East Affiliated to Shanghai University of Medicine and Health Sciences, Shanghai 201306, China; 2. Department of Laboratory Medicine, Shanghai Sixth People’s Hospital East Campus, Shanghai 201306, China
  • Online:2020-07-30 Published:2020-08-06
  • Contact: DU Yuzhen E-mail: duyuzhen2005@163.com

摘要: 背景与目的:斯钙素1(stanniocalcin 1,STC1)与癌症的发展和不良预后相关,但STC1对肺癌细胞转移的影响及其作用机制目前还未完全阐明。探讨STC1对肺癌细胞侵袭和迁移的影响及其可能的内在分子机制。方法:构建STC1过表达的细胞系HLEC-STC1及对照细胞系HLEC-EV,STC1沉默的细胞系A549-STC1 siRNA及对照细胞系A549-EV;采用Transwell和划痕实验检测细胞的侵袭和迁移能力;采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)和蛋白质印迹法(Western blot)检测上皮-间质转化(epithelial-mesenchymal transition,EMT)标志物的mRNA和蛋白质水平变化;并使用细胞免疫荧光技术进一步验证细胞EMT标志物的表达和定位;采用Western blot检测EMT相关信号通路蛋白和转录因子的水平。结果:与对照细胞相比,过表达STC1的HLEC-STC1细胞的侵袭和迁移能力增强,STC1沉默的A549-STC1 siRNA细胞的侵袭和迁移能力减弱(P<0.05);过表达STC1的HLEC-STC1细胞中上皮标志物上皮型钙黏蛋白(E-cadherin)的表达下调,间质标志物神经型钙黏蛋白(N-cadherin)、波形蛋白(vimentin)及肿瘤干细胞标志物上皮细胞黏附分子(epithelial cell adhesion molecule,EpCAM)表达上调(P<0.05);低表达STC1的A549-STC1 siRNA细胞中E-cadherin的表达上调,N-cadherin、vimentin及EpCAM的表达下调(P<0.05);同时,HLEC-STC1细胞中细胞外调节蛋白激酶信号通路(extracellular signal-regulated protein kinase,ERK)及Wnt/β-连环蛋白(β-catenin)信号通路激活相关的磷酸化(p)-ERK和β-catenin的水平升高,转录因子E盒结合锌指蛋白1(zinc finger E-box-binding homeobox 1,ZEB1)和锌指转录因子1(Snail1)的表达水平上调,A549-STC1 siRNA细胞中的结果则相反(P<0.05)。结论:STC1可能通过激活ERK和Wnt/β-catenin信号通路上调转录因子ZEB1和Snail1的表达水平,从而诱导EMT促进肺癌细胞的侵袭和迁移。

关键词: 斯钙素1, 肺癌细胞, 上皮-间质转化, 侵袭, 迁移

Abstract: Background and purpose: Stanniocalcin 1 (STC1) is associated with the development of cancer and poor prognosis. However, the effect of STC1 on lung cancer cell metastasis and its mechanism have not yet been fully clarified. The purpose of this study was to investigate the effect of STC1 on invasion and migration of lung cancer cells and its possible molecular mechanisms. Methods: HLEC-STC1 cell line overexpressing STC1 with the control cell line HLEC-EV and A549-STC1 siRNA cell line silencing STC1 with the control cell line A549-EV were constructed. Transwell and scratch assays were used to detect cell invasion and migration abilities. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot were used to detect the changes of mRNA and protein levels of epithelial-mesenchymal transition (EMT) markers. Cellular immunofluorescence technique was used to further verify the expression and localization of EMT markers. Western blot was used to detect EMT-related signaling pathway proteins and the levels of transcription factors. Results: Compared with the control cells, the invasion and migration abilities of HLEC-STC1 cells overexpressing STC1 were enhanced, while the invasion and migration abilities of STC1-silencing A549-STC1 siRNA cells were weakened (P<0.05).In HLEC-STC1 cells overexpressing STC1,the expression of E-cadherin was down-regulated, while the expression levels of N-cadherin, vimentin and epithelial cell adhesion molecule (EpCAM) were up-regulated (P<0.05). The expression of E-cadherin was up-regulated in A549-STC1 siRNA cells with low expression of STC1. The expression levels of N-cadherin, vimentin and EpCAM were down-regulated (P<0.05). Meanwhile, the expression levels of extracellular signal-regulated protein kinase (ERK) and Wnt/β-catenin signaling pathways in HLEC-STC1 cells were increased, and the expression of transcription factor ZEB1 was elevated. The expression level of Snail1 was up-regulated, and the results were opposite in A549-STC1 siRNA cells (P<0.05). Conclusion: STC1 may up-regulate the expression levels of transcription factors ZEB1 and Snail1 by activating ERK and Wnt/β-catenin signaling pathways, thereby inducing EMT to promote invasion and migration of lung cancer cells.

Key words: Stanniocalcin 1, Lung cancer cells, Epithelial-mesenchymal transition, Invasion, Migration