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Articles in PresS, published online ahead of print April 3, 2002
Am J Physiol Gastrointest Liver Physiol, 10.1152/ajpgi.00495.2001
Submitted on November 19, 2001
Accepted on March 26, 2002
1 Division of Life Sciences, Hallym University, Chunchon, Korea, Republic of
2 Laboratory for Physiological Chemistry, University Medical Center Utrecht, Utrecht, The Netherlands
3 Department of Food and Nutrition, Kyung Hee University, Seoul, Korea, Republic of
4 Division of Applied Life Sciences, Graduate School, Gyeongsang National University, Chinju, Korea, Republic of
5 Department of Pathology, Hallym University, Chunchon, Korea, Republic of
* To whom correspondence should be addressed. E-mail: jyoon{at}hallym.ac.kr.
Previous studies have demonstrated that a commercially available mixture of conjugated linoleic acid (CLA) isomers decreases colon cancer cell growth. The present study was performed to compare the individual potencies of the two main isomers found in the mixture of CLA isomers, e.g. cis-9,trans-11 (c9t11) and trans-10,cis-12 (t10c12), and to assess whether the decreased cell growth is related to changes in secretion of insulin-like growth factor (IGF)-II and/or IGF-binding proteins (IGFBPs) that have been shown to regulate Caco-2 cell proliferation. Cells were incubated in serum-free medium with various concentrations of the individual CLA isomers. T10cc12 CLA decreased cell number in a dose-dependent manner, with a 55 ± 3% reduction in cell number observed within 96 h after the addition of 5 µM t10c12 CLA. The inclusion of t10c12 CLA in culture media induced apoptosis and decreased DNA synthesis. By contrast, c9t11 CLA had no effect. Immunoblot analysis of 24-h, serum-free, conditioned media using a monoclonal anti-IGF-II antibody revealed that Caco-2 cells secreted both mature 7,500 Mr and higher Mr forms of IGF-II. The levels of the higher Mr and the mature form of IGF-II were decreased by treatment with 5 µM t10c12 CLA by 50 ± 3% and 22 ± 2%, respectively. However, c9t11 CLA had no effect. Ligand blot analysis of conditioned medium using 125I-IGF-II revealed that the production of IGFBP-2 was also slightly decreased by t10c12 CLA, whereas c9t11 CLA had no effect. Exogenous IGF-II reversed the growth inhibition and apoptosis induced by t10c12 CLA. These results indicate that inhibition of Caco-2 cell growth by CLA is attributed to the effect of t10c12 CLA, which may be mediated by decreasing IGF-II secretion in Caco-2 cells.
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