Wang Chunyi, Wang Jialin, Liu Hong, Fu Zhongxue
Department of Gastrointestinal Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.
Oncol Rep. 2014 May;31(5):2270-8. doi: 10.3892/or.2014.3057. Epub 2014 Mar 5.
The aim of the present study was to investigate the biological role and molecular mechanism of the deleted in liver cancer-1 (DLC-1) gene in human colon cancer growth and invasion. Recombinant lentiviral vectors encoding the DLC-1 gene were constructed for transfection into the human colon cancer cell line SW480. Real-time quantitative polymerase chain reaction (real-time qPCR) and western blot analysis were employed to evaluate the expression of DLC-1, β-catenin, GSK-3β and c-myc in DLC-1-transfected cells. Moreover, cell proliferation assay, cell colony formation assay, cell cycle analysis, apoptosis analysis and cell migration and invasion assays were performed in order to elucidate the role of DLC-1 in colorectal cancer development and progression. Both real-time qPCR and western blot analyses showed that the DLC-1 gene and protein were overexpressed in the DLC-1-transfected SW480 cells. In addition, the expression of β-catenin and GSK-3β was upregulated and the expression of the c-myc gene was downregulated in the DLC-1-transfected SW480 cells. Furthermore, DLC-1 overexpression inhibited cell proliferation, colony formation, migration and invasion, and induced cell cycle arrest at the G1 phase with subsequent apoptosis. DLC-1 inhibits cell growth and invasion in human colon cancer, functioning as a tumor-suppressor gene, possibly through the regulation of the Wnt/β-catenin signaling pathway.
本研究旨在探讨肝癌缺失基因-1（DLC-1）在人结肠癌生长和侵袭中的生物学作用及分子机制。构建了编码DLC-1基因的重组慢病毒载体，用于转染人结肠癌细胞系SW480。采用实时定量聚合酶链反应（real-time qPCR）和蛋白质免疫印迹分析来评估DLC-1、β-连环蛋白、糖原合成酶激酶-3β（GSK-3β）和c-myc在DLC-1转染细胞中的表达。此外，进行细胞增殖测定、细胞集落形成测定、细胞周期分析、凋亡分析以及细胞迁移和侵袭测定，以阐明DLC-1在结直肠癌发生发展中的作用。实时定量聚合酶链反应和蛋白质免疫印迹分析均显示，DLC-1基因和蛋白在DLC-1转染的SW480细胞中过表达。此外，在DLC-1转染的SW480细胞中，β-连环蛋白和GSK-3β的表达上调，而c-myc基因的表达下调。此外，DLC-1过表达抑制细胞增殖、集落形成、迁移和侵袭，并诱导细胞周期停滞于G1期，随后发生凋亡。DLC-1通过调控Wnt/β-连环蛋白信号通路，作为一种肿瘤抑制基因抑制人结肠癌的细胞生长和侵袭。
