Browse > Article

Study of the Mechanism for the Growth Inhibitory Effects of Conjugated Linoleic Acid on Caco-2 Colon Cancer Cells  

김은지 (한림대학교 실버생물산업기술연구센터)
오윤신 (한림대학교 생명과학부)
이현숙 (한림대학교 생명과학부)
박현서 (경희대학교 식품영양학과)
윤정한 (한림대학교 생명과학부)
Publication Information
Journal of Nutrition and Health / v.36, no.3, 2003 , pp. 270-279 More about this Journal
Abstract
Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of linoleic acid (LA) and exhibits anticarcinogenic activity in a variety of animal models. We have previously observed that CLA inhibited the growth of Caco-2 cells, a human colon adenocarcinoma cell line. The present study was performed to determine whether the growth inhibitory effect of CLA is related to change in secretion of IGF- II and/or IGF-binding proteins (IGFBPs) that have been shown to regulate Caco-2 cell proliferation by an autocrine mechanism. Cells were incubated in serum-free medium with various concentrations of CLA or linoleic acid (LA). Immunoblot analysis of 24-hours, serum-free, conditioned medium using a monoclonal anti-IGF-IIantibody revealed that Caco-2 cells secreted both mature 6,500 Mr and higher Mr forms of pro IGF-II. The levels of pro IGF-II and mature IGF-IIwere decreased by 43 $\pm$ 2% and 53 $\pm$ 6%, respectively by treatment with 50 $\mu$ M CLA. LA slightly increased pro IGF- II levels. Results from Northern blot analysis showed that CLA decreased IGF-II mRNA levels at 50 $\mu$ M concentration suggesting that CLA regulation of IGF-II protein expression occurs partly at the transcriptional level. Ligand blot analysis of conditioned media using 1251-IGF-II revealed that CLA slightly decreased IGFBP-2 levels and increased IGFBP-4 levels. We confirmed our previous results that CLA inhibited cell growth in a dose-dependent manner but LA slightly increased cell growth. Exogenous IGF-II mitigated the growth inhibitory effect of CLA. These results indicate that the growth inhibitory effect of CLA may be at least in part mediated by decreasing IGF-II and IGFBP-2 secretion and increasing IGFBP-4 secretion in Caco-2 cells.
Keywords
conjugated linoleic acid; colon cancer cells; insulin-like growth factor-II; insulin-like growth factor-binding proteins;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Ha YL, Grimm NK, Pariza MW. Newly recognized anticarcinogenic fatty acid: Identification and quantification in natural and processed chesses. J Agr Food Chem 37: 75-81, 1989   DOI
2 Kepler CR, Hirons KP, McNeill JJ, Tove SB. Intermediate and products of the biohydrogenation of linoleic acid by Butyrin-vibrio fibrisolvens. J Biol Chem 241: 1350-1354, 1966
3 Chin SF, Liu W, Storkson JM, Ha YL, Pariza MW. Dietary sources of conjugated dienoic isomers of linoleic acid, a newly recognized class of anticarcinogens. J Food Comp Anal 5: 185-197, 1992   DOI
4 Forgerty A, Ford G, Svomonos D. Octadeca-9, 11-dienoic acid in foodstuffs and in lipids of human blood and breast milk Nutr Rep Int 38: 937-944, 1988
5 Huang YC, Luedecke LO, Shultz TO. Effect of cheddar cheese consumption on plasma conjugated linoleic acid concentrations in men. Nutr Res 14: 373-386, 1994   DOI   ScienceOn
6 Belury MA, Nickel KP, Bird CE, Wu Y. Dietary conjugated linoleic acid modulation of phorbol ester skin turnor promotion. Nutr Cancer 26: 149-157, 1996   DOI   ScienceOn
7 Stewart CE, Rotwein P. Growth, differentiation, and survival: multiple physiological functions for insulin-like growth factors. Physiol Rev 76: 1005-1026, 1996
8 Hwa V, Oh Y, Rosenfeld RG. The insulin-like growth factor-binding protein (IGFBP) superfamily. Endocr Rev 20: 761-787, 1999   DOI   ScienceOn
9 Renehan AG, Painter JE, O'Halloran D, Atkins WS, Potten CS, O'Dwyer ST, Shalet SM. Circulating insulin-like growth factor II and colorectal adenomas. J Clin Endocrinol Metab 85: 3402-3408, 2000   DOI
10 Park HS, Ryu JH, Ha YL, Park JH. Dietary conjugated linoleic acid (CLA) induces apoptosis of colonic mucosa in 1,2-dimethylhydrazine-treated rats: A possible mechanism of the anticarcinogenic effect by CLA. Br J Nutr 86: 549-555, 2001   DOI   ScienceOn
11 O'Shea M, Stanton C, Devery R. Antioxidant enzyme defence responses of human MCF-7 and SW480 cancer cells to conjugated linoleic acid. Anticancer Res 19: 1953-1959, 1999
12 Zarrilli R, Pignata S, Romano M, Gravina A, Casola S, Bruni CB, Acquaviva AM. Expression of insulin-like growth factor (IGF)-II and IGF-I receptor during proliferation and differentiation of CaCo-2 human colon carcinoma cells. Cell Growth Differ 5:1085-1091, 1994
13 Oguchi S, Walker WA, Sanderson IR. Profile of IGF-binding proteins secreted by intestinal epithelila cells changes with differentiation. Am J Physiol 267: G843-G850, 1994
14 Hoeflich A, Yang Y, Huber S, Rascher W, Keopf G, Blum WF, Heinz-Erian P, Kolb HJ, Kiess W. Expression of IGFBP-2, -3, and -4 mRNA during differentiation of Caco-2 epithelial cells. Am J Physiol 271: E922-E931, 1996
15 Park JH, Corkins MR, Vanderhoof JA, Caruso NM, Hrbek MJ, Schaffer BS, Slentz DH, McCusker RH, MacDonald RG. Expression of insulin-like growth factor-II and insulin-like growth factor binding proteins during Caco-2 cell proliferation and differentiation. J Cell Physiol 166: 396-406, 1996   DOI
16 Miraki-Moud F, Ooi B, Sidawy A, Korman L. Role of insulin-like growth factor binding protein-2 (IGFBP-2) stimulates proliferation and decreases apoptosis in Caco-2 colon cancer cells. Endocr Soc 83rd Annu Meet Progr Abstr, 196, 2001
17 Kim EJ, Kim WY, Kang YH, Ha YL, Bach LA, Park JHY. 2000. Inhibition of Caco-2 cell proliferation by (n-3) fatty acids: possible mediation by increased secretion of insulin-like growth factor binding protein-6. Nutr Res 20: 1409-1421   DOI   ScienceOn
18 Jones JI, Clemmons DR. Insulin-like growth factors and their binding proteins: biological actions. Endocrine Rev 16: 3-34, 1995
19 Buckbinder L, Talbott R, velasco-Miguel S, Tokenaka I, Faha B, Seizinger BR, Kley N. Induction of the growth inhibitor IGF-binding protein-3 by p53. Nature 377: 646-649, 1995   DOI   ScienceOn
20 Kim EJ, Holthuizen PE, Park HS, Ha YL, Jung KC, Park JHY. Trans-lO,cis-12-conjugated linoleic acid inhibits Caco-2 colon cancer cell growth. Am J Physiol Gastrointest Liver Physiol 283:G357-G367, 2002
21 Kim EJ, Schaffer BS, Kang YH, Macdonald RG, Park JH. Decreased production of insulin-like growth factor-binding protein (IGFBP)-6 by transfection of colon cancer cells with an antisense IGFBP-6 cDNA construct leads to stimulation of cell proliferation. J Gastroenterol Hepatol 17: 563-570, 2002   DOI   ScienceOn
22 Oh YS, Kim EJ, Lee SG, Chung C-K, Kang I-J, Shin HK, Park JHY. Induction of apoptosis cell death and depression of Bcl-2 protein levels by trans-10, cis-12 comjugated linoleic acid in human prostate cancer. J Korean Soc Food Sci Nutr 31: 1126-1133, 2002   DOI
23 Knekt P, Jarvinen R, Seppanen R, Pukkala E, Aromaa A. Intake of dairy products and the risk of breast cancer. Br J Cancer 73:687-691, 1996   DOI   ScienceOn
24 Zhang L, Zhou W, Velculescu VE, Kern SE, Hurban RH, Hamilton SR, Vogelstein B, Kinzler KW. Gene expression profiles in normal and cancer cells. Science 276: 1268-1272, 1997   DOI   ScienceOn
25 Ip C. Review of the effects of trans fatty acids, oleic acid, n-3 polyunsaturated fatty acids, and conjugated linoleic acid on mammary carcinogenesis in animals. Am J Clin Nutr 66: 1523S-1529S, 1997
26 Oh YS, Kim EJ, Kim JW, Kim WK, Lee HS, Yoon JH. The effect of conjugated linoleic acid and its isomers on the proliferation of prostate TSU-Prl cancer cells. Korean J Nutrition 35: 192-200, 2002
27 McCarty MF. Activation of PPAR gamma may mediated a portion of the anticancer activity of conjugated linoleic acid. Medical Hypothesis 55: 187-188, 2000   DOI   ScienceOn
28 Ip C, Carter CA, Ip MM. Requirement of essential fatty acid for mammary tumorigenesis in the rat. Cancer Res 45: 1997-2001, 1985
29 Jansen M, van Schaik FM, van Tol H, Van den Brande JL, Sussenbach JS. Nucleotide sequences of cDNAs encoding precursors of human insulin-like growth factor II (IGF-II) and an IGF-II variant. FEBS Lett 179: 243,1985   DOI   ScienceOn
30 Underwood LE. Nutritional regulation of IGF-I and IGFBPs. J Pediatr Endocrinol and metab 9: 302-312, 1996
31 Ip C, Briggs SP, Haegele AD, Thompson HJ, Storkson J, Scimeca JA. The conjugated linoleic acid in mammary cancer prevention is independent of the level or type of fat in the diet. Carcinogenesis 17: 1045-1050, 1996   DOI   ScienceOn
32 Mishra L, Bass B, Ooi B, Sidawy A, Korman L. Role of insulin-like growth factor-I (IGF-I) receptor, IGF-I, and IGF binding protein-2 in human colorectal cancer. Growth Horm IGF Res 8: 473-479, 1998   DOI   ScienceOn
33 Ha YL, Grimm NK, Pariza MW. Anticarcinogens from fried ground beef: heat-altered derivatives of linoleic acid. Carcinogenesis 8: 1881-1887, 1987   DOI   ScienceOn
34 Beserga R. Oncogenes and the strategy of growth factors. Cell 79: 927-930, 1995   DOI   ScienceOn
35 Denizot F, Lang R. 1986. Rapid colorimetric assay for cell growth and survival modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunological Methods 89: 271-277   DOI   ScienceOn
36 Erickson KL. Is there a relation between dietary linoleic acid and cancer of the breast, colon, or prostate? Am J Clin Nutr 68:5-7, 1998
37 Cesano A, Visonneau S, Scimeca JA, Kritchevsky D, Santoli D. Opposite effect of linoleic acid and conjugated linoleic acid on human prostatic cancer in SCID mice. Anticancer Res 18: 1429-1434, 1998
38 Park HS. Mechanism of cancer prevention and other physiological function by conjugated linoleic acid. Korean J Nutrition 33: 556-565, 2000
39 Yu Y, Correll PH, Vanden Heuvel JP. Conjugated linoleic acid decreases production of pro-inflammatory products in macrophages: evidence for a PPAR gamma-dependent mechanism. Biochim Biophys Acta 1581: 89-99, 2002   DOI   ScienceOn
40 Kim EJ, Kang YH, Schaffer BS, Bach LA, MacDonald RG, Park JH. Inhibition of Caco-2 cell proliferation by all-trans retinoic acid: role of insulin-like growth factor binding protein-6. J Cell Physiol 190: 92-100, 2002   DOI   ScienceOn
41 Visonneau S, Cesano A, Tepper SA, Scimeca J, Santoli D, Kritchevsky D. Conjugated linoleic acid suppresses the growth of human brest adenocarcinoma cells in SCID mice. Anticancer Res 17: 969-974, 1997   DOI   ScienceOn
42 Oh Y. IGF-independent regulation of breast cancer growth by IGF binding proteins. Breast Cancer Res Treat 47: 283-293, 1998   DOI   ScienceOn
43 Shultz TD, Chew BP, Seaman WR. Differential stimulatory and inhibitory responses of human MCF-7 breast cancer cells to linoleic acid and conjugated linoleic acid in culture. Anticancer Res 12: 2143-2145, 1992
44 Schonberg S, Krokan HE. The inhibitory effect of conjugated linoleic acid derivatives (CLA) of linoleic acid on the growth of human tumor cell lines is in part due to increased lipid peroxidation. Anticancer Res 15: 1241-1246, 1995
45 Kim EJ, Cho HJ, Kim SJ, Kang Y-H, Ha Y-L, Park JHY. Effect of conjugated linoleic acid on the proliferation of the human colon cancer cell line, HT-29. Korean J Nutrition 34: 894-904, 2001
46 Kim EJ, Jun J-G, Park HS, Kim SM, Ha YL, Park JHY. Conjugated linoleic acid (CLA) inhibits growth of Caco-2 colon cancer cells: possible mediation by oleamide. Anticancer Res 22: 2193-2198, 2002
47 Liew C, Schutt HAJ, Chin SF, Pariza MW, Dashwood RH. Protection of conjugated linoleic acids against 2-amino-3-methylimidazo[4,5-f] quinoline-induced colon carcinogenesis in the F344 Rat: A study of inhibitory mechanisms. Carcinogenesis 16:3037-3043, 1995   DOI   ScienceOn
48 Chomczynski P. 1993. A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques 15: 536-537