Top

International Journal of Colorectal Disease

Published in:

01-04-2007 | Original Article

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

Authors: Olorunseun O. Ogunwobi, Ian L. P. Beales

Published in: International Journal of Colorectal Disease | Issue 4/2007

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.

Tuynman JB, Peppelenbosch MP, Richel DJ (2004) COX-2 inhibition as a tool to treat and prevent colorectal cancer. Crit Rev Oncol Hematol 52:81–101PubMed

Giovannucci E, Ascherio A, Rimm EB, Colditz GA, Stampfer MJ, Willett WC (1995) Physical activity, obesity, and risk for colon cancer and adenoma in men. Ann Intern Med 122:327–334PubMed

Pan SY, Johnson KC, Ugnat AM, Wen SW, Mao Y (2004) Association of obesity and cancer risk in Canada. Am J Epidemiol 159: 259–268PubMedCrossRef

Considine RV, Sinha MK, Heiman ML, Kriauciunas A, Stephens TW, Nyce MR, Ohannesian JP, Marco CC, McKee LJ, Bauer TL et al (1996) Serum immunoreactive-leptin concentrations in normal-weight and obese humans. N Engl J Med 334:292–295PubMedCrossRef

Stattin P, Lukanova A, Biessy C, Soderberg S, Palmqvist R, Kaaks R, Olsson T, Jellum E (2004) Obesity and colon cancer: does leptin provide a link? Int J Cancer 109:149–152PubMedCrossRef

Attoub S, Noe V, Pirola L, Bruyneel E, Chastre E, Mareel M, Wymann MP, Gespach C (2000) Leptin promotes invasiveness of kidney and colonic epithelial cells via phosphoinositide 3-kinase-, rho-, and rac-dependent signaling pathways. FASEB J 14:2329–2338PubMedCrossRef

Hardwick JC, van Den Brink GR, Offerhaus GJ, van Deventer SJ, Peppelenbosch MP (2001) Leptin is a growth factor for colonic epithelial cells. Gastroenterology 121:79–90PubMedCrossRef

Myers MG Jr (2004) Leptin receptor signaling and the regulation of mammalian physiology. Recent Prog Horm Res 59:287–304PubMedCrossRef

Saxena NK, Titus MA, Ding X, Floyd J, Srinivasan S, Sitaraman SV, Anania FA (2004) Leptin as a novel profibrogenic cytokine in hepatic stellate cells: mitogenesis and inhibition of apoptosis mediated by extracellular regulated kinase (Erk) and Akt phosphorylation. FASEB J 18:1612–1614PubMed

10.

Kim GS, Hong JS, Kim SW, Koh JM, An CS, Choi JY, Cheng SL (2003) Leptin induces apoptosis via ERK/cPLA2/cytochrome c pathway in human bone marrow stromal cells. J Biol Chem 278:21920–21929PubMedCrossRef

11.

Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J, Greenberg ME (1999) Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96:857–868PubMedCrossRef

12.

Thompson JE, Thompson CB (2004) Putting the rap on Akt. J Clin Oncol 22:4217–4226PubMedCrossRef

13.

Somasundar P, Frankenberry KA, Skinner H, Vedula G, McFadden DW, Riggs D, Jackson B, Vangilder R, Hileman SM, Vona-Davis LC (2004) Prostate cancer cell proliferation is influenced by leptin. J Surg Res 118:71–82PubMedCrossRef

14.

Stepan VM, Sawada M, Todisco A, Dickinson CJ (1999) Glycine-extended gastrin exerts growth-promoting effects on human colon cancer cells. Mol Med 5:147–159PubMed

15.

Ogunwobi O, Beales I (2005) Glycine-extended gastrin stimulates proliferation and inhibits apoptosis in colon cancer cells via multiple pathways (abstract). Gut 54(Suppl 11):A57

16.

Onuma M, Bub JD, Rummel TL, Iwamoto Y (2003) Prostate cancer cell–adipocyte interaction: leptin mediates androgen-independent prostate cancer cell proliferation through c-Jun NH2-terminal kinase. J Biol Chem 278:42660–42667PubMedCrossRef

17.

Sakai I, Kraft AS (1997) The kinase domain of Jak2 mediates induction of bcl-2 and delays cell death in hematopoietic cells. J Biol Chem 272:12350–12358PubMedCrossRef

18.

Kloek C, Haq AK, Dunn SL, Lavery HJ, Banks AS, Myers MG Jr (2002) Regulation of Jak kinases by intracellular leptin receptor sequences. J Biol Chem 277:41547–41555PubMedCrossRef

19.

Pai R, Lin C, Tran T, Tarnawski A (2005) Leptin activates STAT and ERK2 pathways and induces gastric cancer cell proliferation. Biochem Biophys Res Commun 331:984–992PubMedCrossRef

20.

Vaisse C, Halaas JL, Horvath CM, Darnell JE Jr, Stoffel M, Friedman JM (1996) Leptin activation of Stat3 in the hypothalamus of wild-type and ob/ob mice but not db/db mice. Nat Genet 14:95–97PubMedCrossRef

21.

Ma XT, Wang S, Ye YJ, Du RY, Cui ZR, Somsouk M (2004) Constitutive activation of Stat3 signaling pathway in human colorectal carcinoma. World J Gastroenterol 10:1569–1573PubMed

22.

Steinbach G, Lynch PM, Phillips RK, Wallace MH, Hawk E, Gordon GB, Wakabayashi N, Saunders B, Shen Y, Fujimura T et al (2000) The effect of celecoxib, a cyclooxygenase-2 inhibitor, in familial adenomatous polyposis. N Engl J Med 342:1946–1952PubMedCrossRef

23.

Corpet DE, Pierre F (2005) How good are rodent models of carcinogenesis in predicting efficacy in humans? A systematic review and meta-analysis of colon chemoprevention in rats, mice and men. Eur J Cancer 41:1911–1922PubMedCrossRef

24.

Maier TJ, Schilling K, Schmidt R, Geisslinger G, Grosch S (2004) Cyclooxygenase-2 (COX-2)-dependent and -independent anticarcinogenic effects of celecoxib in human colon carcinoma cells. Biochem Pharmacol 67:1469–1478PubMedCrossRef

25.

Beales I, Calam J (1997) Stimulation of IL-8 production in human gastric epithelial cells by Helicobacter pylori, IL-1β and TNF-α requires tyrosine kinase activity, but not protein kinase C. Cytokine 9:514–520PubMedCrossRef

26.

Beales IL (2004) Gastrin and interleukin-1beta stimulate growth factor secretion from cultured rabbit gastric parietal cells. Life Sci 75:2983–2995PubMedCrossRef

27.

Micoud F, Mandrand B, Malcus-Vocanson C (2001) Comparison of several techniques for the detection of apoptotic astrocytes in vitro. Cell Prolif 34:99–113PubMedCrossRef

28.

Ogunwobi OO, Beales IL (2006) Glycine-extended gastrin stimulates proliferation and inhibits apoptosis in colon cancer cells via cyclo-oxygenase-independent pathways. Regul Pept 134:1–8PubMedCrossRef

29.

Ostlund RE Jr, Yang JW, Klein S, Gingerich R (1996) Relation between plasma leptin concentration and body fat, gender, diet, age, and metabolic covariates. J Clin Endocrinol Metab 81:3909–3913PubMedCrossRef

30.

Zimmet P, Hodge A, Nicolson M, Staten M, de Courten M, Moore J, Morawiecki A, Lubina J, Collier G, Alberti G et al (1996) Serum leptin concentration, obesity, and insulin resistance in Western Samoans: cross sectional study. BMJ 313:965–969PubMed

31.

Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, Greenberg ME (1997) Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91:231–241PubMedCrossRef

32.

Skov S, Klausen P, Claesson MH (1997) Ligation of major histocompatibility complex (MHC) class I molecules on human T cells induces cell death through PI-3 kinase-induced c-Jun NH2-terminal kinase activity: a novel apoptotic pathway distinct from Fas-induced apoptosis. J Cell Biol 139:1523–1531PubMedCrossRef

33.

Bjorbaek C, Uotani S, da Silva B, Flier JS (1997) Divergent signaling capacities of the long and short isoforms of the leptin receptor. J Biol Chem 272:32686–32695PubMedCrossRef

34.

Arico S, Pattingre S, Bauvy C, Gane P, Barbat A, Codogno P, Ogier-Denis E (2002) Celecoxib induces apoptosis by inhibiting 3-phosphoinositide-dependent protein kinase-1 activity in the human colon cancer HT-29 cell line. J Biol Chem 277:27613–27621PubMedCrossRef

35.

Kumar NV, Bernstein LR (2001) Ten ERK-related proteins in three distinct classes associate with AP-1 proteins and/or AP-1 DNA. J Biol Chem 276:32362–32372PubMedCrossRef

36.

Zhong SP, Ma WY, Quealy JA, Zhang Y, Dong Z (2001) Organ-specific distribution of AP-1 in AP-1 luciferase transgenic mice during the maturation process. Am J Physiol Regul Integr Comp Physiol 280:R376–R381PubMed

37.

Shaulian E, Karin M (2001) AP-1 in cell proliferation and survival. Oncogene 20:2390–2400PubMedCrossRef

38.

Ashida R, Tominaga K, Sasaki E, Watanabe T, Fujiwara Y, Oshitani N, Higuchi K, Mitsuyama S, Iwao H, Arakawa T (2005) AP-1 and colorectal cancer. Inflammopharmacology 13:113–125PubMedCrossRef

39.

Rivat C, Rodrigues S, Bruyneel E, Pietu G, Robert A, Redeuilh G, Bracke M, Gespach C, Attoub S (2005) Implication of STAT3 signaling in human colonic cancer cells during intestinal trefoil factor 3 (TFF3)—and vascular endothelial growth factor-mediated cellular invasion and tumor growth. Cancer Res 65:195–202PubMed

40.

Bromberg J (2002) Stat proteins and oncogenesis. J Clin Invest 109:1139–1142PubMedCrossRef

41.

Rouet-Benzineb P, Aparicio T, Guilmeau S, Pouzet C, Descatoire V, Buyse M, Bado A (2004) Leptin counteracts sodium butyrate_induced apoptosis in human colon cancer HT-29 cells via NF-kappaB signaling. J Biol Chem 279:16495–16502PubMedCrossRef

Title: 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
Authors: Olorunseun O. Ogunwobi
Ian L. P. Beales
Publication date: 01-04-2007
Publisher: Springer-Verlag
Published in: International Journal of Colorectal Disease / Issue 4/2007
Print ISSN: 0179-1958
Electronic ISSN: 1432-1262
DOI: https://doi.org/10.1007/s00384-006-0181-y

Keynote webinar | Spotlight on medication adherence

Springer Medicine

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

Abstract

Background and aims

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background and aims

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 4/2007

Investigation of association of the DLG5 gene with phenotypes of inflammatory bowel disease in the British population

Acceptance of flexible sigmoidoscopy as a screening examination for colorectal cancer in an outpatient clinic

NOD2/CARD15 gene polymorphisms are not associated with collagenous colitis

The posterior approach for low retrorectal tumors in adults

Capecitabine-induced pancolitis

Increased detection rate and potential prognostic impact of disseminated tumor cells in patients undergoing endorectal ultrasound for rectal cancer