Original Article
Copyright ©2014 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Mar 28, 2014; 20(12): 3287-3300
Published online Mar 28, 2014. doi: 10.3748/wjg.v20.i12.3287
Cortactin expression confers a more malignant phenotype to gastric cancer SGC-7901 cells
Jun Wei, Zhong-Xin Zhao, Yang Li, Zhu-Qing Zhou, Tian-Geng You
Jun Wei, Zhong-Xin Zhao, Yang Li, Zhu-Qing Zhou, Tian-Geng You, Department of Gastroenterological Surgery, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China
Author contributions: You TG and Zhao ZX designed the study; Wei J, Li Y and Zhou ZQ performed the experiments; Wei J and Zhou ZQ analyzed the data; Wei J wrote the paper; all of the authors contributed to the preparation of the manuscript.
Supported by Grants from the Project of Shanghai Science and Technology Commission of China, No. 10411968400
Correspondence to: Tian-Geng You, PhD, Department of Gastroenterological Surgery, Shanghai East Hospital, Tongji University School of Medicine, Jimo Rd 150, Shanghai 200120, China. tiangengyou2003@163.com
Telephone: +86-21-65982142 Fax: +86-21-58798999
Received: November 15, 2013
Revised: February 10, 2014
Accepted: March 6, 2014
Published online: March 28, 2014
Processing time: 132 Days and 1.2 Hours
Abstract

AIM: To study the effects of cortactin on the tumor biology of SGC-7901 cells and identify the mechanism involved in the process.

METHODS: Cell lines in which cortactin was stably overexpressed or knocked down as well as the respective control cell lines were established by standard molecular methods. The effects of cortactin on the proliferation, migration and invasion capacity of SGC-7901 cells were assessed by the MTT assay, colony formation, flow cytometry, transwell migration and matrigel invasion. Nude mouse models were also used to assess the role of cortactin in the growth and metastasis of SGC-7901 cells in vivo. Western blotting analysis was performed to detect the expression of epidermal growth factor receptor (EGFR) and downstream molecules.

RESULTS: Cell lines in which cortactin was stably overexpressed or knocked down as well as control cell lines were successfully established and designated as LV5-cortactin-SGC, LV5-SGC, LV3-shRNA-SGC and LV3-SGC. Cortactin overexpression promoted SGC-7901 cell migration (340.7 ±12.6 vs 229.1 ± 23.2, P < 0.01) and invasion (71.6 ± 5.2 vs 48.4 ± 3.6, P < 0.01). Cortactin downregulation impaired SGC-7901 cell migration (136.2 ± 19.8 vs 225 ± 17) and invasion (29.2 ± 5.2 vs 49.6 ± 3.8, P < 0.01). The results from the MTT and colony formation assays results indicated increased LV5-cortactin-SGC cell proliferation and decreased LV3-shRNA-SGC cell proliferation compared to the control cells. Flow cytometry analysis demonstrated that cortactin overexpression promoted the proliferation index of SGC-7901 cells, and the results were reversed when cortactin was downregulated. Mouse tumor models confirmed that cortactin expression increased SGC-7901 cell proliferation and metastasis in vivo. Western blotting analysis revealed that cortactin elevated EGFR expression and activated the downstream molecules.

CONCLUSION: Cortactin expression promoted the migration, invasion and proliferation of SGC-7901 cells both in vivo and in vitro. The EGFR signaling pathway is mechanistically involved.

Keywords: Gastric cancer; Cortactin; Epidermal growth factor receptor; Invasion; Metastasis; Proliferation

Core tip: Cortactin is an actin-related protein 2/3 complex-activating and filamentous (F)-actin-binding protein that is implicated in tumor cell motility and metastasis. Clinical studies have shown that cortactin overexpression is often associated with the clinicopathological parameters and poor prognosis in a variety of cancers, including gastric cancer. In this study, the effects of cortactin on gastric cancer progression were investigated. The results showed that cortactin expression promoted SGC-7901 cell migration, invasion and proliferation both in vitro and in vivo. The epidermal growth factor receptor signaling pathway is mechanistically involved. Cortactin may serve as a novel therapeutic target of gastric cancer.