Blockade of long chain fatty acid oxidation by non-steroidal anti-inflammatory drugs may contribute to inhibition of proliferation of human colorectal carcinoma cell lines
Introduction
Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit colorectal carcinoma (CRC) growth. For example, indomethacin [1], piroxicam [2]and sulindac [3]inhibit the development of chemically-induced CRC in rodents and sulindac also reduces the size and number of colorectal polyps in patients with familial adenomatous polyposis [4]. In addition, several epidemiological studies have revealed that NSAIDs, and in particular aspirin, reduce the risk of CRC and other cancers of the gastrointestinal tract by approximately 50% 5, 6. In the presence of NSAIDs cells of diverse origin [7]are reversibly arrested in the G1 phase, with no impairment of viability [8]. The anti-inflammatory effects of NSAIDs result from inhibition of cyclooxygenases [9]and selective antagonists have indicated that the inducible isozyme cyclooxygenase-2 is also one of the targets for the inhibitory effects of NSAIDs on CRC growth in vivo [10].
Several lines of evidence suggest that some of the growth-inhibitory effects of NSAIDs in vitro are mediated by different targets. For example, antagonists selective for cyclooxygenase-2 have no effect on the growth of CRC cell lines [11]. On the other hand, the sulfone derivative of the NSAID sulindac does not inhibit cyclooxygenases in vitro, but is an effective inhibitor of the growth of CRC cell lines [12]. Evidence is also accumulating that sulindac and its sulfone derivative can induce apoptosis both in CRC cell lines [13]and in the colorectal epithelium [14]. However, the observation that aspirin inhibits proliferation 12, 15but does not induce apoptosis [15]in a CRC cell line indicates that other inhibitory pathways must also be present in vitro and may contribute to growth inhibition in vivo. Because some NSAIDs have previously been reported to inhibit long chain fatty acid oxidation in liver mitochondria 16, 17, 18, we wished to determine firstly whether or not NSAIDs inhibited long chain fatty acid oxidation in CRC cells and secondly whether such inhibition contributed to the inhibitory effects of NSAIDs on the growth of CRC cells. Therefore, we measured the potencies of NSAIDs as inhibitors of long chain fatty acid oxidation in human CRC cell lines and of the growth of the human CRC cell line LIM 1215. Comparison of the IC50 (IC50, concentration required for 50% inhibition) values obtained suggests that one or more of the enzymes of the long chain fatty acid oxidation pathway may be targets for the anti-proliferative effects of NSAIDs.
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Materials
Palmitic acid, fatty acid-free fraction V bovine serum albumin and NSAIDs were from Sigma (St. Louis, MO, USA), with the exception of sulindac sulfide and sulfone which were a generous gift from Merck, Sharp and Dohme (West Point, PA). [9,10(n)-3H]palmitic acid and [3H]sodium acetate were from Amersham (Little Chalfont, Bucks, UK) and DuPont NEN (Boston, MA, USA), respectively. AG1-8X anion exchange resin was from Bio-Rad (North Ryde, Australia). The human CRC cell lines LIM 1215 [19]and LIM
Inhibition of long chain fatty acid oxidation by NSAIDs in CRC cells
All of the NSAIDs tested inhibited oxidation of the long chain fatty acid palmitate by the CRC cell line LIM 1215 (Fig. 1, Table 1). Similar inhibition was also observed in the CRC cell line LIM 1899 (data not shown). This is the first report of inhibition of long chain fatty acid oxidation by NSAIDs in cells derived from the colonic epithelium. Previous reports of effects of NSAIDs on long chain fatty acid oxidation have been limited to inhibition of palmitate oxidation by pirprofen [16],
Acknowledgements
We thank Dr Bob Whitehead for many helpful discussions. This work was supported in part by grants #920527 and #960182 from the National Health and Medical Research Council of Australia.
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Present address: Faculté de Pharmacie, Université de Montpellier 1, 34060 Montpellier Cedex, France.