Abstract
Hyperinsulinemia and type II diabetes are associated with an increased risk of developing colorectal tumors. We found previously that in intestinal cells, insulin or insulin-like growth factor-1 stimulates c-Myc and cyclin D1 protein expression through both Akt-dependent and Akt-independent mechanisms. The effect of Akt is attributed to the stimulation of c-Myc translation by mammalian target of rapamycin. However, Akt-independent stimulation was, associated with an increase in β-catenin (β-cat) in the nucleus and an increased association between β-cat and T-cell factor binding sites on the c-Myc promoter, detected by chromatin immunoprecipitation. In this study, we show that insulin stimulated the phosphorylation/activation of p-21-activated protein kinase-1 (PAK-1) in an Akt-independent manner in vitro and in an in vivo hyperinsulinemic mouse model. Significantly, shRNA (small hairpin RNA)-mediated PAK-1 knockdown attenuated both basal and insulin-stimulated c-Myc and cyclin D1 expression, associated with a marked reduction in extracellular signal-regulated kinase activation and β-cat phosphorylation at Ser675. Furthermore, PAK-1 silencing led to a complete blockade of insulin-stimulated β-cat binding to the c-Myc promoter and cellular growth. Finally, inhibition of MEK, a downstream target of PAK-1, blocked insulin-stimulated nuclear β-cat accumulation and c-Myc expression. Our observations suggest that PAK-1 serves as an important linker between insulin and Wnt signaling pathways.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
References
Aberle H, Bauer A, Stappert J, Kispert A, Kemler R . (1997). beta-Catenin is a target for the ubiquitin-proteasome pathway. Embo J 16: 3797–3804.
Adam L, Vadlamudi R, Mandal M, Chernoff J, Kumar R . (2000). Regulation of microfilament reorganization and invasiveness of breast cancer cells by kinase dead p21-activated kinase-1. J Biol Chem 275: 12041–12050.
Aguilera O, Fraga MF, Ballestar E, Paz MF, Herranz M, Espada J et al. (2006). Epigenetic inactivation of the Wnt antagonist DICKKOPF-1 (DKK-1) gene in human colorectal cancer. Oncogene 25: 4116–4121.
Akinmade D, Talukder AH, Zhang Y, Luo WM, Kumar R, Hamburger AW . (2008). Phosphorylation of the ErbB3 binding protein Ebp1 by p21-activated kinase 1 in breast cancer cells. Br J Cancer 98: 1132–1140.
Alessi DR, Cuenda A, Cohen P, Dudley DT, Saltiel AR . (1995). PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J Biol Chem 270: 27489–27494.
Arias-Romero LE, Chernoff J . (2008). A tale of two Paks. Biol Cell 100: 97–108.
Behrens J, von Kries JP, Kuhl M, Bruhn L, Wedlich D, Grosschedl R et al. (1996). Functional interaction of beta-catenin with the transcription factor LEF-1. Nature 382: 638–642.
Bienz M, Clevers H . (2000). Linking colorectal cancer to Wnt signaling. Cell 103: 311–320.
Bustin S . (2001). Protein shuttles, IGF-I and colorectal cancer. Trends Mol Med 7: 9.
Carter JH, Douglass LE, Deddens JA, Colligan BM, Bhatt TR, Pemberton JO et al. (2004). Pak-1 expression increases with progression of colorectal carcinomas to metastasis. Clin Cancer Res 10: 3448–3456.
Chang CK, Ulrich CM . (2003). Hyperinsulinaemia and hyperglycaemia: possible risk factors of colorectal cancer among diabetic patients. Diabetologia 46: 595–607.
Ching YP, Leong VY, Lee MF, Xu HT, Jin DY, Ng IO . (2007). P21-activated protein kinase is overexpressed in hepatocellular carcinoma and enhances cancer metastasis involving c-Jun NH2-terminal kinase activation and paxillin phosphorylation. Cancer Res 67: 3601–3608.
Clements WM, Lowy AM, Groden J . (2003). Adenomatous polyposis coli/beta-catenin interaction and downstream targets: altered gene expression in gastrointestinal tumors. Clin Colorectal Cancer 3: 113–120.
Clevers H . (2004). Wnt breakers in colon cancer. Cancer Cell 5: 5–6.
Coles LC, Shaw PE . (2002). PAK1 primes MEK1 for phosphorylation by Raf-1 kinase during cross-cascade activation of the ERK pathway. Oncogene 21: 2236–2244.
Desbois-Mouthon C, Cadoret A, Blivet-Van Eggelpoel MJ, Bertrand F, Cherqui G, Perret C et al. (2001). Insulin and IGF-1 stimulate the beta-catenin pathway through two signalling cascades involving GSK-3beta inhibition and Ras activation. Oncogene 20: 252–259.
Dong YF, Soung do Y, Schwarz EM, O'Keefe RJ, Drissi H . (2006). Wnt induction of chondrocyte hypertrophy through the Runx2 transcription factor. J Cell Physiol 208: 77–86.
Dudley DT, Pang L, Decker SJ, Bridges AJ, Saltiel AR . (1995). A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci USA 92: 7686–7689.
Eblen ST, Slack JK, Weber MJ, Catling AD . (2002). Rac-PAK signaling stimulates extracellular signal-regulated kinase (ERK) activation by regulating formation of MEK1-ERK complexes. Mol Cell Biol 22: 6023–6033.
Elwing JE, Gao F, Davidson NO, Early DS . (2006). Type 2 diabetes mellitus: the impact on colorectal adenoma risk in women. Am J Gastroenterol 101: 1866–1871.
Giovannucci E . (2001). Insulin, insulin-like growth factors and colon cancer: a review of the evidence. J Nutr 131: 3109S–3120S.
Giovannucci E . (2007). Metabolic syndrome, hyperinsulinemia, and colon cancer: a review. Am J Clin Nutr 86: s836–s842.
He H, Shulkes A, Baldwin GS . (2008). PAK1 interacts with beta-catenin and is required for the regulation of the beta-catenin signalling pathway by gastrins. Biochim Biophys Acta 1783: 1943–1954.
He TC, Sparks AB, Rago C, Hermeking H, Zawel L, da Costa LT et al. (1998). Identification of c-MYC as a target of the APC pathway. Science 281: 1509–1512.
Hino S, Tanji C, Nakayama KI, Kikuchi A . (2005). Phosphorylation of {beta}-catenin by cyclic AMP-dependent protein kinase stabilizes {beta}-catenin through inhibition of its ubiquitination. Mol Cell Biol 25: 9063–9072.
Hinoi T, Yamamoto H, Kishida M, Takada S, Kishida S, Kikuchi A . (2000). Complex formation of adenomatous polyposis coli gene product and axin facilitates glycogen synthase kinase-3 beta-dependent phosphorylation of beta-catenin and down-regulates beta-catenin. J Biol Chem 275: 34399–34406.
Ilyas M, Tomlinson IP, Rowan A, Pignatelli M, Bodmer WF . (1997). Beta-catenin mutations in cell lines established from human colorectal cancers. Proc Natl Acad Sci USA 94: 10330–10334.
Inoki K, Ouyang H, Zhu T, Lindvall C, Wang Y, Zhang X et al. (2006). TSC2 integrates Wnt and energy signals via a coordinated phosphorylation by AMPK and GSK3 to regulate cell growth. Cell 126: 955–968.
Jin S, Zhuo Y, Guo W, Field J . (2005). p21-activated Kinase 1 (Pak1)-dependent phosphorylation of Raf-1 regulates its mitochondrial localization, phosphorylation of BAD, and Bcl-2 association. J Biol Chem 280: 24698–24705.
Jin T, Fantus IG, Sun J . (2008). Wnt and beyond Wnt: multiple mechanisms control the transcriptional property of beta-catenin. Cell Signal 20: 1697–1704.
King AJ, Sun H, Diaz B, Barnard D, Miao W, Bagrodia S et al. (1998). The protein kinase Pak3 positively regulates Raf-1 activity through phosphorylation of serine 338. Nature 396: 180–183.
King CC, Gardiner EM, Zenke FT, Bohl BP, Newton AC, Hemmings BA et al. (2000). p21-activated kinase (PAK1) is phosphorylated and activated by 3-phosphoinositide-dependent kinase-1 (PDK1). J Biol Chem 275: 41201–41209.
Kinzler KW, Vogelstein B . (1996). Lessons from hereditary colorectal cancer. Cell 87: 159–170.
Knaus UG, Bokoch GM . (1998). The p21Rac/Cdc42-activated kinases (PAKs). Int J Biochem Cell Biol 30: 857–862.
Lewin HS . (2008). Diabetes mellitus publication patterns, 1984-2005. J Med Libr Assoc 96: 155–158.
Li J, Mizukami Y, Zhang X, Jo WS, Chung DC . (2005). Oncogenic K-ras stimulates Wnt signaling in colon cancer through inhibition of GSK-3beta. Gastroenterology 128: 1907–1918.
Li W, Chong H, Guan KL . (2001). Function of the Rho family GTPases in Ras-stimulated Raf activation. J Biol Chem 276: 34728–34737.
Liu C, Li Y, Semenov M, Han C, Baeg GH, Tan Y et al. (2002). Control of beta-catenin phosphorylation/degradation by a dual-kinase mechanism. Cell 108: 837–847.
Liu Z, Habener JF . (2008). Glucagon-like peptide-1 activation of TCF7L2-dependent Wnt signaling enhances pancreatic beta cell proliferation. J Biol Chem 283: 8723–8735.
Ma J, Giovannucci E, Pollak M, Leavitt A, Tao Y, Gaziano JM et al. (2004). A prospective study of plasma C-peptide and colorectal cancer risk in men. J Natl Cancer Inst 96: 546–553.
Mann B, Gelos M, Siedow A, Hanski ML, Gratchev A, Ilyas M et al. (1999). Target genes of beta-catenin-T cell-factor/lymphoid-enhancer-factor signaling in human colorectal carcinomas. Proc Natl Acad Sci USA 96: 1603–1608.
Molenaar M, van de Wetering M, Oosterwegel M, Peterson-Maduro J, Godsave S, Korinek V et al. (1996). XTcf-3 transcription factor mediates beta-catenin-induced axis formation in Xenopus embryos. Cell 86: 391–399.
Olnes MI, Kurl RN . (1994). Isolation of nuclear extracts from fragile cells: a simplified procedure applied to thymocytes. Biotechniques 17: 828–829.
Park ER, Eblen ST, Catling AD . (2007). MEK1 activation by PAK: a novel mechanism. Cell Signal 19: 1488–1496.
Playford MP, Bicknell D, Bodmer WF, Macaulay VM . (2000). Insulin-like growth factor 1 regulates the location, stability, and transcriptional activity of beta-catenin. Proc Natl Acad Sci USA 97: 12103–12108.
Powell SM, Zilz N, Beazer-Barclay Y, Bryan TM, Hamilton SR, Thibodeau SN et al. (1992). APC mutations occur early during colorectal tumorigenesis. Nature 359: 235–237.
Qiao M, Shapiro P, Kumar R, Passaniti A . (2004). Insulin-like growth factor-1 regulates endogenous RUNX2 activity in endothelial cells through a phosphatidylinositol 3-kinase/ERK-dependent and Akt-independent signaling pathway. J Biol Chem 279: 42709–42718.
Rochat A, Fernandez A, Vandromme M, Moles JP, Bouschet T, Carnac G et al. (2004). Insulin and wnt1 pathways cooperate to induce reserve cell activation in differentiation and myotube hypertrophy. Mol Biol Cell 15: 4544–4555.
Sells MA, Knaus UG, Bagrodia S, Ambrose DM, Bokoch GM, Chernoff J . (1997). Human p21-activated kinase (Pak1) regulates actin organization in mammalian cells. Curr Biol 7: 202–210.
Slack-Davis JK, Eblen ST, Zecevic M, Boerner SA, Tarcsafalvi A, Diaz HB et al. (2003). PAK1 phosphorylation of MEK1 regulates fibronectin-stimulated MAPK activation. J Cell Biol 162: 281–291.
Smith SD, Jaffer ZM, Chernoff J, Ridley AJ . (2008). PAK1-mediated activation of ERK1/2 regulates lamellipodial dynamics. J Cell Sci 121: 3729–3736.
Sun J, Jin T . (2008). Both Wnt and mTOR signaling pathways are involved in insulin-stimulated proto-oncogene expression in intestinal cells. Cell Signal 20: 219–229.
Sundberg-Smith LJ, Doherty JT, Mack CP, Taylor JM . (2005). Adhesion stimulates direct PAK1/ERK2 association and leads to ERK-dependent PAK1 Thr212 phosphorylation. J Biol Chem 280: 2055–2064.
Tang Y, Chen Z, Ambrose D, Liu J, Gibbs JB, Chernoff J et al. (1997). Kinase-deficient Pak1 mutants inhibit Ras transformation of Rat-1 fibroblasts. Mol Cell Biol 17: 4454–4464.
Tang Y, Marwaha S, Rutkowski JL, Tennekoon GI, Phillips PC, Field J . (1998). A role for Pak protein kinases in Schwann cell transformation. Proc Natl Acad Sci USA 95: 5139–5144.
Tang Y, Zhou H, Chen A, Pittman RN, Field J . (2000). The Akt proto-oncogene links Ras to Pak and cell survival signals. J Biol Chem 275: 9106–9109.
Taurin S, Sandbo N, Qin Y, Browning D, Dulin NO . (2006). Phosphorylation of beta-catenin by cyclic AMP-dependent protein kinase. J Biol Chem 281: 9971–9976.
Tetsu O, McCormick F . (1999). Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature 398: 422–426.
Tran TT, Medline A, Bruce WR . (1996). Insulin promotion of colon tumors in rats. Cancer Epidemiol Biomarkers Prev 5: 1013–1015.
Tsakiridis T, Taha C, Grinstein S, Klip A . (1996). Insulin activates a p21-activated kinase in muscle cells via phosphatidylinositol 3-kinase. J Biol Chem 271: 19664–19667.
Wang P, Branch DR, Bali M, Schultz GA, Goss PE, Jin T . (2003). The POU homeodomain protein OCT3 as a potential transcriptional activator for fibroblast growth factor-4 (FGF-4) in human breast cancer cells. Biochem J 375: 199–205.
Yi F, Brubaker PL, Jin T . (2005). TCF-4 mediates cell type-specific regulation of proglucagon gene expression by beta-catenin and glycogen synthase kinase-3beta. J Biol Chem 280: 1457–1464.
Yi F, Sun J, Lim GE, Fantus IG, Brubaker PL, Jin T . (2008). Cross talk between the insulin and Wnt signaling pathways: evidence from intestinal endocrine L cells. Endocrinology 149: 2341–2351.
Yoshii S, Tanaka M, Otsuki Y, Fujiyama T, Kataoka H, Arai H et al. (2001). Involvement of alpha-PAK-interacting exchange factor in the PAK1-c-Jun NH(2)-terminal kinase 1 activation and apoptosis induced by benzo[a]pyrene. Mol Cell Biol 21: 6796–6807.
Zhang Q, Adiseshaiah P, Kalvakolanu DV, Reddy SP . (2006). A Phosphatidylinositol 3-kinase-regulated Akt-independent signaling promotes cigarette smoke-induced FRA-1 expression. J Biol Chem 281: 10174–10181.
Zhou GL, Field J . (2004). Targeting PAK etk. Cancer Biol Ther 3: 102–103.
Zhou GL, Zhuo Y, King CC, Fryer BH, Bokoch GM, Field J . (2003). Akt phosphorylation of serine 21 on Pak1 modulates Nck binding and cell migration. Mol Cell Biol 23: 8058–8069.
Acknowledgements
We thank Dr Jeffrey Field (University of Pennsylvania) for providing the dominant negative PAK-1 (K299R), constitutively active PAK-1 (T423E) and control plasmids. JS is a recipient of Banting and Best Diabetes Centre (BBDC) Novo Nordisk graduate studentship (2004–2008). This study was supported by an operating grant from Canadian Institutes of Health Research to TJ (MOP-89987) and a Translational Acceleration Research Grant from the Canadian Breast Cancer Research Alliance/CIHR (# 016512) to IGF.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)
Supplementary information
Rights and permissions
About this article
Cite this article
Sun, J., Khalid, S., Rozakis-Adcock, M. et al. P-21-activated protein kinase-1 functions as a linker between insulin and Wnt signaling pathways in the intestine. Oncogene 28, 3132–3144 (2009). https://doi.org/10.1038/onc.2009.167
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2009.167
Keywords
This article is cited by
-
P21-activated kinase 2-mediated β-catenin signaling promotes cancer stemness and osimertinib resistance in EGFR-mutant non-small-cell lung cancer
Oncogene (2022)
-
Behavioral Risk Factors and Risk of Early-Onset Colorectal Cancer: Review of the Mechanistic and Observational Evidence
Current Colorectal Cancer Reports (2021)
-
Group I Paks are essential for epithelial- mesenchymal transition in an Apc-driven model of colorectal cancer
Nature Communications (2018)
-
Curcumin represses mouse 3T3-L1 cell adipogenic differentiation via inhibiting miR-17-5p and stimulating the Wnt signalling pathway effector Tcf7l2
Cell Death & Disease (2017)
-
Dysregulated glycolysis as an oncogenic event
Cellular and Molecular Life Sciences (2015)