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International Journal of Colorectal Disease

, Volume 22, Issue 4, pp 401–409 | Cite as

The anti-apoptotic and growth stimulatory actions of leptin in human colon cancer cells involves activation of JNK mitogen activated protein kinase, JAK2 and PI3 kinase/Akt

  • Olorunseun O. Ogunwobi
  • Ian L. P. BealesEmail author
Original Article

Abstract

Background and aims

Obesity is a major risk factor for the development of colon cancer. Secretion of the hormone leptin from adipocytes is increased in obesity, and serum levels are proportional to body fat mass. Serum leptin levels are an independent risk factor for colon cancer. Leptin receptors are expressed in normal, premalignant and malignant colonic epithelia. We have investigated the effects of leptin on proliferation and apoptosis of colonic cancer cells and the early signalling events involved.

Methods

Proliferation of HT-29 colon cancer cells in response to leptin was assessed by 3-[4, 5-dimethylthiazol-2-y-l]-2, 5-diphenyltetrazolium bromide (MTT) assay, and apoptosis was quantified by enzyme-linked immunosorbent assay (ELISA) for intracellular nucleosomes. Signalling pathways involved were determined by using specific inhibitors, quantification of phosphorylated active intermediates and ELISA of active nuclear-translocated transcription factors.

Results

Leptin stimulated HT-29 cell proliferation and inhibited both serum-starvation and celecoxib-induced apoptosis. The proliferative and anti-apoptotic effects of leptin were abolished by inhibition of JAK2 with AG490, phosphatidylinositol 3′-kinase (PI3 kinase) with LY294002 and c-Jun NH2-terminal kinase (JNK) with SP600125. Stimulation of HT-29 cells with leptin increased phosphorylation of JAK2, Akt and JNK. Activation of JAK2 was upstream of PI3 kinase/Akt but not of JNK. Activation of JAK2 was followed by activation and nuclear translocation of STAT3 and JNK activation led to increased activator protein 1 (AP-1) transcriptional activity.

Conclusions

Leptin stimulates proliferation and inhibits apoptosis in human colon cancer cells and may be an important factor in the increased incidence of colon cancer in obesity. This effect involves JAK2, PI3 kinase and JNK and activation of the oncogenic transcription factors signal transducer and activator of transcription (STAT)3 and AP-1.

Keywords

Leptin Apoptosis Proliferation Colon cancer Janus tyrosine kinase 2 Akt c-Jun NH2-terminal kinase Activator protein 1 STAT3 protein Celecoxib 

Abbreviations

AP-1

activator protein 1

COX

cyclo-oxygenase

DMEM

Dulbecco’s modified Eagle medium

DMSO

dimethyl sulphoxide

ELISA

enzyme-linked immunosorbent assay

FBS

foetal bovine serum

JAK

Janus tyrosine kinase

JNK

c-Jun NH2-terminal kinase

MAP

mitogen-activated protein kinase

MTT

3-[4, 5-dimethylthiazol-2-y-l]-2, 5-diphenyltetrazolium bromide

PBS

phosphate-buffered saline (pH 7.4)

PI3 kinase

phosphatidylinositol 3′-kinase

STAT

signal transducer and activator of transcription

Notes

Acknowledgements

This work was funded by the Norfolk and Norwich University Hospital Bicentenary Trust in the form of a research studentship to Dr. Ogunwobi. Further financial support was provided by The Royal Society, The Peel Medical Research Trust, The Mason Medical Research Foundation, The Institute of Biomedical Science, the Norfolk and Norwich University Hospital Inflammatory Bowel Disease Research Fund and National Health Service (NHS) Research and Development. Part of this work was presented in abstract form to the British Society of Gastroenterology, March 2005.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Gastroenterology DepartmentNorfolk and Norwich University HospitalNorwichUK
  2. 2.Gastroenterology Research Unit, School of Medicine, Health Policy and PracticeUniversity of East AngliaNorwichUK

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