Overexpression of Contactin 1 promotes growth, migration and invasion in Hs578T breast cancer cells

Abstract

Background: Contactin1 (CNTN1) has been shown to play an important role in the invasion and metastasis of several tumors; however, the role of CNTN1 in breast cancer has not been fully studied. The purpose of this study is to investigate the role of CNTN1 in regulating tumor growth, migration and invasion in breast cancer.

Results: To investigate its function, CNTN1 was expressed in Hs578T cells. CNTN1 expression was confirmed by western blot, immunohistochemistry and real-time RT-PCR. The effect of CNTN1 overexpression on proliferation, migration and invasion of Hs578T breast cancer cells was assessed in vitro and in vivo. Our results showed that CNTN1 overexpression promoted Hs578T cell proliferation, cell cycle progression, colony formation, invasion and migration. Notably, overexpression of CNTN1 in Hs578T cells enhanced the growth of mouse xenograft tumors.

Conclusions: CNTN1 promotes growth, metastasis and invasion of Hs578T breast cancer cell line. Thus, therapies targeting CNTN1 may prove efficacious for breast cancer. However, further investigation is required to understand the mechanism by which CNTN1 influences proliferation, metastasis and invasion in breast cancer.

Keywords: Breast cancer; CNTN1; Hs578T; Invasion; Migration; Proliferation.

Fig. 1

Expression of CNTN1 in breast cancer cell lines. a Expression of CNTN1 in MCF7-ADR, MDA-MB-468, MCF7 and Hs578T cells was assayed by western blot. CNTN1 was the least expression compared with controls (^*P<0.05) (b). Western blot. Bands were quantified by densitometry and CNTN1 was normalized to B-actin levels. c Real-time RT-PCR. Expression of CNTN1 mRNA was the least expression compared with controls (^*P<0.05)

Fig. 2

Expression of CNTN1 in Hs578T cells. a Immunocytochemical staining 48 h after transfection, b Real-time RT-PCR: Expression of CNTN1 was significantly increased in CNTN1-transfected cells (P<0.05). c Western blot. CNTN1 was overexpressed compared with controls (P<0.05), d Western blotting quantitation

Fig. 3

Exogenous CNTN1 induces cell proliferation and growth of Hs578T cells. a Cell cycle analysis of CNTN1 transfected cells and untransfected cells; percentage of cells in each phase of the cell cycle are indicated. The experiments were performed in triplicate and results are represented as mean ± standard error. b Colony formation assay indicates that CNTN1 overexpression led to an increase in colony number; c MTT assay of CNTN1 overexpressing cells: compared to controls, OD was significantly higher in CNTN1-transfected group (*P<0.05); d Representative bar graph

Fig. 4

CNTN1 overexpression induces migration and invasion of Hs578T cells. Migration (a) and invasion (b) assays showed a higher fraction of pEGFP-N1-CNTN1 cells migrated and invaded, than untreated and pEGFP-N1 cells (c) Representative bar graph (P<0.05)

Fig. 5

CNTN1 overexpression promotes growth of Hs578T cells in mouse xenograft model. a Representative images of tumors collected from mice at day 21. b Overexpression of CNTN1 promotes growth Hs578T tumors in vivo (n = 6/group). Data are expressed as mean ± standard error. *P < 0.05; pEGFP-N1-CNTN1 vs. untreated or pEGFP-N1 cell groups. c Immunohistochemical staining of tumor specimens. Data are represented as mean ± S.D. (* p < 0.05)