Antitumorigenic Effect of a High Protein Diet in Mouse Skin

  • Tak, Ka-Hee (Department of Food Science and Nutrition, Catholic University of Daegu) ;
  • Kim, Eun-Jung (Department of Food Science and Nutrition, Catholic University of Daegu)
  • Received : 2011.08.29
  • Accepted : 2011.10.10
  • Published : 2011.12.31


The recent increase of colon, breast, and prostate cancer incidence in Korea has been attributed to a diet pattern change to a more Western style, in which the foods eaten are higher in protein and fat. Whether high protein intake itself stimulates tumor cell growth and exacerbates disease status has been investigated, however, many epidemiological studies have inconsistent results between meat intake and the risk of certain cancers. These inconsistent results are partly because of the difficulty of studying the effects of just the meat intake. Other factors, such as overall meal context, could not be completely excluded in the study. To address the question of whether high protein itself is independently associated with carcinogenesis, we initiated ICR mice with 200 nmol ($50{\mu}g$) 7,12-dimethylbenz[a]anthracene (DMBA) and fed animals either a normal diet (ND, 14% casein) or a high protein diet (HPD, 50% casein) for 15 weeks with 12-O-tetradecanoylphorbol-13-acetate (TPA) promotion in two-stage skin carcinogenesis protocol. There was no significant difference between ND and HPD group in food intake and body weight throughout the experiment. However, tumor multiplicity of the HPD group was decreased by 75.5% compared to that of the ND group. In addition, HPD inhibited skin hyperplasia and epidermal cell proliferation. Western analyses with whole skin lysates showed that HPD inhibited TPA-induced Akt (S473), S6K (T389), 4E-BP1 (Thr 37/46) and Erk1/2 (Thr202/Tyr204) phosphorylation as well as COX-2 expression. Taken together, these data suggest that a high protein diet has an anticarcinogenic effect by inhibiting the TPA-induced Akt signaling pathway.


Supported by : National Research Foundation of Korea (NRF)


  1. Doll R, Peto R. 1981. The causes of cancer: quantitative estimates of avoidable risks of cancer in the United States today. J Natl Cancer Inst 66: 1191-1308.
  2. Allen NE, Key TJ, Appleby PN, Travis RC, Roddam AW, Tjønneland A, Johnsen NF, Overvad K, Linseisen J, Rohrmann S, Boeing H, Pischon T, Bueno-de-Mesquita HB, Kiemeney L, Tagliabue G, Palli D, Vineis P, Tumino R, Trichopoulou A, Kassapa C, Trichopoulos D, Ardanaz E, Larrañaga N, Tormo MJ, González CA, Quirós JR, Sánchez MJ, Bingham S, Khaw KT, Manjer J, Berglund G, Stattin P, Hallmans G, Slimani N, Ferrari P, Rinaldi S, Riboli E. 2008. Animal foods, protein, calcium and prostate cancer risk: the European Prospective Investigation into Cancer and Nutrition. Br J Cancer 98: 1574-1581.
  3. Lee JE, Spiegelman D, Hunter DJ, Albanes D, Bernstein L, van den Brandt PA, Buring JE, Cho E, English DR, Freudenheim JL, Giles GG, Graham S, Horn-Ross PL, Håkansson N, Leitzmann MF, Männistö S, McCullough ML, Miller AB, Parker AS, Rohan TE, Schatzkin A, Schouten LJ, Sweeney C, Willett WC, Wolk A, Zhang SM, Smith-Warner SA. 2008. Fat, protein, and meat consumption and renal cell cancer risk: a pooled analysis of 13 prospective studies. J Natl Cancer Inst 100: 1695-1706.
  4. Hill M. 2002. Meat, cancer and dietary advice to the public. Eur J Clin Nutr 56: S36-S41.
  5. Kim E. 2009. Mechanisms of amino acid sensing in mTOR signaling pathway. Nutr Res Prac 3: 64-71.
  6. Slaga TJ, Fischer SM, Weeks CE, Klein-Szanto AJ, Reiners J. 1982. Studies on the mechanisms involved in multistage carcinogenesis in mouse skin. J Cell Biochem 18: 99-119.
  7. Slaga TJ, Fischer SM, Nelson K, Gleason GL. 1980. Studies on the mechanism of skin tumor promotion: evidence for several stages in promotion. Proc Natl Acad Sci USA 77: 3659-3663.
  8. DiGiovanni J. 1992. Multistage carcinogenesis in mouse skin. Pharmacol Ther 54: 63-128.
  9. Slaga TJ, Klein-Szanto AJ, Fischer SM, Weeks CE, Nelson K, Major S. 1980. Studies on mechanism of action of anti- tumor-promoting agents: their specificity in two-stage promotion. Proc Natl Acad Sci USA 77: 2251-2254.
  10. Boutwell RK, Verma AK, Ashendel CL, Astrup E. 1982. Mouse skin: a useful model system for studying the mechanism of chemical carcinogenesis. Carcinog Compr Surv 7: 1-12.
  11. Brown K, Buchmann A, Balmain A. 1990. Carcinogen-induced mutations in the mouse c-Ha-ras gene provide evidence of multiple pathways for tumor progression. Proc Natl Acad Sci USA 87: 538-542.
  12. Slaga TJ. 1983. Overview of tumor promotion in animals. Environ Health Persp 50: 3-14.
  13. Winberg LD, Budunova IV, Warren BS, Lyer RP, Slaga TJ. 1995. Mechanisms of skin tumor promotion and progression. CRC Press, Inc., Boca Raton, FL, USA. p 113-120.
  14. Niedel JE, Kuhn LJ, Vandenbark GR. 1983. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci USA 80: 36-40.
  15. Castagna M, Takai Y, Kaibuchi K, Sano K, Kikkawa U, Nishizuka Y. 1982. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem 257: 7847-7851.
  16. Marks F, Furstenberger G. 2000. Cancer chemoprevention through interruption of multistage carcinogenesis. The lessons learnt by comparing mouse skin carcinogenesis and human large bowel cancer. Eur J Cancer 36: 314-329.
  17. Tang Q, Gonzales M, Inoue H, Bowden GT. 2001. Roles of Akt and glycogen synthase kinase 3beta in the ultraviolet B induction of cyclooxygenase-2 transcription in human keratinocytes. Cancer Res 61: 4329-4332.
  18. Stephens L, Anderson K, Stokoe D, Erdjument-Bromage H, Painter GF, Holmes AB, Gaffney PR, Reese CB, Mc- Cormick F, Tempst P, Coadwell J, Hawkins PT. 1998. Protein kinase B kinases that mediate phosphatidylinositol 3,4,5-trisphosphate-dependent activation of protein kinase B. Science 279: 710-714.
  19. Chan TO, Rittenhouse SE, Tsichlis PN. 1999. AKT/PKB and other D3 phosphoinositide-regulated kinases: kinase activation by phosphoinositide-dependent phosphorylation. Annu Rev Biochem 68: 965-1014.
  20. Kim E. 2010. Insulin resistance at the crossroads of metabolic syndrome: systemic analysis using microarrays. Biotechnol J 5: 919-929.
  21. Testa JR, Tsichlis PN. 2005. AKT signaling in normal and malignant cells. Oncogene 24: 7391-7393.
  22. Lu J, Rho O, Wilker E, Beltran L, Digiovanni J. 2007. Activation of epidermal akt by diverse mouse skin tumor promoters. Mol Cancer Res 5: 1342-1352.
  23. Alexander DD, Cushing CA. 2011. Red meat and colorectal cancer: a critical summary of prospective epidemiologic studies. Obes Rev 12: e472-e493.
  24. Alexander DD, Mink PJ, Cushing CA, Sceurman B. 2010. A review and meta-analysis of prospective studies of red and processed meat intake and prostate cancer. Nutr J 9: 50.
  25. Alexander DD, Morimoto LM, Mink PJ, Cushing CA. 2010. A review and meta-analysis of red and processed meat consumption and breast cancer. Nutr Res Rev 23: 349-365.
  26. McAfee AJ, McSorley EM, Cuskelly GJ, Moss BW, Wallace JM, Bonham MP, Fearon AM. 2010. Red meat consumption: an overview of the risks and benefits. Meat Sci 84: 1-13.
  27. Ferguson LR. 2010. Meat and cancer. Meat Sci 84: 308-313.
  28. Santarelli RL, Pierre F, Corpet DE. 2008. Processed meat and colorectal cancer: a review of epidemiologic and experimental evidence. Nutr Cancer 60: 131-144.
  29. Ho VW, Leung K, Hsu A, Luk B, Lai J, Shen SY, Minchinton AI, Waterhouse D, Bally MB, Lin W, Nelson BH, Sly LM, Krystal G. 2011. A low carbohydrate, high protein diet slows tumor growth and prevents cancer initiation. Cancer Res 71: 4484-4493.
  30. Rose ML, Madren J, Bunzendahl H, Thurman RG. 1999. Dietary glycine inhibits the growth of B16 melanoma tumors in mice. Carcinogenesis 20: 793-798.
  31. Z'graggen K, Warshaw, AL, Werner J, Graeme-Cook F, Jimenez RE, Fernández-del Castillo C. 2001. Promoting effect of a high-fat/high-protein diet in DMBA-induced ductal pancreatic cancer in rats. Ann Surg 233: 688-695.
  32. Segrelles C, Ruiz S, Perez P, Murga C, Santos M, Budunova IV, Martinez J, Larcher F, Slaga TJ, Gutkind JS, Jorcano JL, Paramio JM. 2002. Functional roles of Akt signaling in mouse skin tumorigenesis. Oncogene 21: 53-64.
  33. Segrelles C, Lu J, Hammann B, Santos M, Moral M, Cascallana JL, Lara MF, Rho O, Carbajal S, Traag J, Beltran L, Martinez-Cruz AB, Garcia-Escudero R, Lorz C, Ruiz S, Bravo A, Paramio JM, DiGiovanni J. 2007. Deregulated activity of Akt in epithelial basal cells induces spontaneous tumors and heightened sensitivity to skin carcinogenesis. Cancer Res 67: 10879-10888.
  34. Hara K, Maruki Y, Long X, Yoshino K, Oshino N, Hidayat S, Tokunaga C, Avruch J, Yonezawa K. 2002. Raptor, a binding partner of target of rapamycin (TOR), mediates TOR action. Cell 110: 177-189.
  35. Kim DH, Sarbassov DD, Ali SM, King JE, Latek RR, Erdjument-Bromage H, Tempst P, Sabatini DM. 2002. mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the cell growth machinery. Cell 110: 163-175.
  36. Sarbassov DD, Ali SM, Kim DH, Guertin DA, Latek RR, Erdjument-Bromage H, Tempst P, Sabatini DM. 2004. Rictor, a novel binding partner of mTOR, defines a rapamycin- insensitive and raptor-independent pathway that regulates the cytoskeleton. Curr Biol 14: 1296-1302.
  37. Fingar DC, Salama S, Tsou C, Harlow E, Blenis J. 2002. Mammalian cell size is controlled by mTOR and its downstream targets S6K1 and 4EBP1/eIF4E. Genes Dev 16: 1472-1487.
  38. Lee CH, Inoki K, Guan KL. 2007. mTOR pathway as a target in tissue hypertrophy. Annu Rev Pharmacol Toxicol 47: 443-467.
  39. Kim J, Kundu M, Viollet B, Guan KL. 2011. AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol 13: 132-141.
  40. Inoki K, Ouyang H, Li Y, Guan KL. 2005. Signaling by target of rapamycin proteins in cell growth control. Microbiol Mol Biol Rev 69: 79-100.
  41. Yang Q, Guan KL. 2007. Expanding mTOR signaling. Cell Res 17: 666-681.
  42. Corradetti MN, Guan KL. 2006. Upstream of the mammalian target of rapamycin: do all roads pass through mTOR? Oncogene 25: 6347-6360.
  43. Laplante M, Sabatini DM. 2009. mTOR signaling at a glance. J Cell Sci 122: 3589-3594.
  44. Kim J, Guan KL. 2011. Amino acid signaling in TOR activation. Ann Rev Biochem 80: 1001-1032.
  45. Checkley L, Rho O, Moore T, Hursting S, DiGiovanni J. 2011. Rapamycin is a potent inhibitor of skin tumor promotion by 12-o-tetradecanoylphorbol-13-acetate. Cancer Prev Res 4: 1011-1020.
  46. Hunot S, Vila M, Teismann P, Davis RJ, Hirsch EC, Przedborski S, Rakic P, Flavell RA. 2004. JNK-mediated induction of cyclooxygenase 2 is required for neurodegeneration in a mouse model of Parkinson's disease. Proc Natl Acad Sci USA 101: 665-670.
  47. Guan Z, Buckman SY, Pentland AP, Templeton DJ, Morrison AR. 1998. Induction of cyclooxygenase-2 by the activated MEKK1 $\rightarrow$ SEK1/MKK4 $\rightarrow$ p38 mitogen-activated protein kinase pathway. J Biol Chem 273: 12901-12908.
  48. Chen W, Tang Q, Gonzales MS, Bowden GT. 2001. Role of p38 MAP kinases and ERK in mediating ultraviolet-B induced cyclooxygenase-2 gene expression in human keratinocytes. Oncogene 20: 3921-3926.
  49. Jang BC, Kim DH, Park JW, Kwon TK, Kim SP, Song DK, Park JG, Bae JH, Mun KC, Baek WK, Suh MH, Hla T, Suh SI. 2004. Induction of cyclooxygenase-2 in macrophages by catalase: role of NF-kappaB and PI3K signaling pathways. Biochem Biophys Res Commun 316: 398-406.
  50. Van Dross RT, Hong X, Pelling JC. 2005. Inhibition of TPA-induced cyclooxygenase-2 (COX-2) expression by apigenin through downregulation of Akt signal transduction in human keratinocytes. Mol Carcinogen 44: 83-91.