Elsevier

Life Sciences

Volume 88, Issues 25–26, 20 June 2011, Pages 1108-1112
Life Sciences

Effects of adrenaline in human colon adenocarcinoma HT-29 cells

https://doi.org/10.1016/j.lfs.2011.04.007Get rights and content

Abstract

Aims

Stress has been implicated in the development of cancers. Adrenaline levels are increased in response to stress. The effects of adrenaline on colon cancer are largely unknown. The aims of the study are to determine the effects of adrenaline in human colon adenocarcinoma HT-29 cells and the possible underlying mechanisms involved.

Main methods

The effect of adrenaline on HT-29 cell proliferation was determined by [3H] thymidine incorporation assay. Expression of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) were detected by Western blot. Matrix metalloproteinase-9 (MMP-9) activity and prostaglandin E2 (PGE2) release were determined by zymography and enzyme immunoassay, respectively.

Key findings

Adrenaline stimulated HT-29 cell proliferation. This was accompanied by the enhanced expression of COX-2 and VEGF in HT-29 cells. Adrenaline also upregulated MMP-9 activity and PGE2 release. Adrenaline stimulated HT-29 cell proliferation which was reversed by COX-2 inhibitor sc-236. COX-2 inhibitor also reverted the action of adrenaline on VEGF expression and MMP-9 activity. Further study was performed to determine the involvement of β-adrenoceptors. The stimulatory action of adrenaline on colon cancer growth was blocked by atenolol and ICI 118,551, a β1- and β2-selective antagonist, respectively. This signified the role of β-adrenoceptors in this process. In addition, both antagonists also abrogated the stimulating actions of adrenaline on COX-2, VEGF expression, MMP-9 activity and PGE2 release in HT-29 cells.

Significance

These results suggest that adrenaline stimulates cell proliferation of HT-29 cells via both β1- and β2-adrenoceptors by a COX-2 dependent pathway.

Introduction

Stress has been linked to the increased incidence and development of cancers (Antoni et al., 2006, Reiche et al., 2004). Adrenaline and noradrenaline are the most important catecholamines released during stress. Recent studies suggested that chronic stress promotes tumor growth and angiogenesis in vivo and catecholamines increase angiogenic cytokines in various cancer cells in vitro (Lutgendorf et al., 2003, Thaker et al., 2006, Wong et al., 2009, Yang et al., 2006).

Colon cancer is the leading cause of cancers and cancer deaths worldwide (Jemal et al., 2010). Expression of β-adrenoceptors have been identified on colon cancer cells (Odore et al., 2003, Wu et al., 2005). Previous studies have shown that activation of β-adrenoceptors has been implicated in the arachidonic acid pathway especially cyclooxygenase-2 (COX-2) expression in relation to pulmonary and pancreatic cancer growth (Schuller et al., 1999, Weddle et al., 2001). The protective effects of nonsteroidal antiinflammatory drugs and COX-2 inhibitors in the treatment and prevention of colon cancers suggest the key role of COX-2 in the pathogenesis of colon cancer (Rice et al., 2003, Steinbach et al., 2000). Stimulation of β-adrenoceptors by catecholamines have also been shown to play a role in the progression of ovarian cancer and are associated with the production of vascular endothelial growth factor (VEGF) (Lutgendorf et al., 2003). In another study, activation of β-adrenoceptors by catecholamines increased the expression of matrix metalloproteinases (MMPs) in nasopharyngeal carcinoma tumor cells (Yang et al., 2006). VEGF is expressed by most human cancers including colon cancer (Brown et al., 1993, Folkman, 1990). The expression profiles of MMPs in different human cancers have been suggested to be associated with progression of the disease. Among them, MMP-9 has been implicated in colon cancer (Fingleton, 2006, Mook et al., 2004). All these findings indicate that there is a possible link between β-adrenoceptors and some of the carcinogenic mediators including COX-2, VEGF and MMP-9 in the development of colon cancer.

In the present study, we aimed to determine the actions of adrenaline on HT-29 human colon cancer cells and further investigate the possible underlying mechanisms involved. This study might shed light on understanding the carcinogenic mechanism of colon cancer and possibly open up a new therapeutic avenue for the prevention and treatment of colon cancer, especially related to stress.

Section snippets

Reagents

Adrenaline, atenolol (β1-selective antagonists), ICI 118,551 (β2-selective antagonists) and antibody for β-actin were purchased from Sigma (St. Louis, MO, USA). sc-236 (4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide) was purchased from Pharmacia (Peapack, NJ, USA). Antibodies for COX-2 and VEGF were purchased from Santa Cruz Biotechnology (Santa Cruz, CA¸ USA). Prostaglandin E2 (PGE2) enzyme immunoassay kit was obtained from R&D Systems (Minneapolis, MN, USA).

Cell culture and viability assays

The

Adrenaline increased HT-29 cell proliferation

Adrenaline stimulated HT-29 colon cancer cell proliferation in a dose-dependent manner (Fig. 1). Adrenaline at the concentration of 10 μM significantly increased HT-29 cell proliferation by 35% when compared to the control group.

Adrenaline upregulated COX-2, VEGF expression, MMP-9 activity and PGE2 release in HT-29 cells

Further study was performed to determine the possible associated pathways in the promotion of HT-29 cell growth by adrenaline. It was found that adrenaline treatment significantly increased COX-2, VEGF expression and PGE2 release (Fig. 2A and C). Adrenaline also

Discussion

Growing evidence have suggested that stress mediators are implicated in cancer growth and progression. However, the effects of adrenaline on colon cancer growth and the underlying molecular and cellular events are largely unknown. In the present study, we show that adrenaline could stimulate HT-29 cell proliferation. Adrenaline also increased COX-2, VEGF expression, MMP-9 activity and PGE2 release in HT-29 cells. The increase of cell proliferation by adrenaline was reversed respectively by

Conclusion

In this study, we showed that adrenaline stimulates HT-29 cell proliferation via both β1- and β2-adrenoceptors by a COX-2 dependent pathway. These findings are important in understanding the pathogenesis of colon cancer, especially related to stress.

Conflict of interest statement

The authors declare that there are no conflicts of interest.

Acknowledgments

The study was supported by the Committee on Research and Conference Grant of the University of Hong Kong, the Direct Grant for Research from the Chinese University of Hong Kong and the Hong Kong Research Grants Council (CUHK 7499/05M).

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