Abstract
Rats subcutaneously implanted with AH109A hepatoma cells show hyperlipidemia with high concentrations of serum triglyceride and nonesterified fatty acid, suppression of lipoprotein lipase (LPL), and elevation of hormone-sensitive lipase (HSL) activities during the growth of the hepatoma. Supplementation of the diet with sulfur amino acids such as l-methionine (Met) and l-cystine (Cys) improved hyperlipidemia by restoring LPL and HSL activities. In the present study, we have attempted to examine the effects of sulfur amino acids on the activity and mRNA level of LPL and the activity of HSL using 3T3-L1 cells, which are known to differentiate to adipocytes. The adipocytes were incubated with various concentrations of Met, Cys or l-cysteine (CysH) in the absence or presence of tumor necrosis factor-α (TNF-α). LPL activity was suppressed by TNF-α. In the absence of TNF-α, Met, Cys and CysH did not change the LPL activity. In the presence of TNF-α, Met and Cys significantly increased the LPL activity, and Met also enhanced the LPL mRNA level. HSL activity was also suppressed by TNF-α. In the absence of TNF-α, Met enhanced the HSL activity. In the presence of TNF-α, Met, Cys and CysH suppressed the HSL activity. Sulfur amino acids such as Met, Cys and CysH affected the LPL activity, mRNA level, and HSL activity in 3T3-L1 adipocytes. Some of these effects of sulfur amino acids were different between LPL and HSL, between the absence and the presence of TNF-α, and between 3T3-L1 adipocytes and the adipose tissue from rats.
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10616-010-9282-0/MediaObjects/10616_2010_9282_Fig1_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10616-010-9282-0/MediaObjects/10616_2010_9282_Fig2_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10616-010-9282-0/MediaObjects/10616_2010_9282_Fig3_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10616-010-9282-0/MediaObjects/10616_2010_9282_Fig4_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10616-010-9282-0/MediaObjects/10616_2010_9282_Fig5_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10616-010-9282-0/MediaObjects/10616_2010_9282_Fig6_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10616-010-9282-0/MediaObjects/10616_2010_9282_Fig7_HTML.gif)
Similar content being viewed by others
References
Barclay M, Skipski VP, Terebus-Kekish O, Greene EM, Kaufman RJ, Stock CC (1970) Effects of cancer upon high-density and other lipoproteins. Cancer Res 30:2420–2430
Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein using principles of protein–dye binding. Anal Biochem 72:248–254
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol–chloroform extraction. Anal Biochem 162:156–159
Fort P, Marty L, Piechaczyk M, El Sabrouty S, Dani C, Jeanteur P, Blanchard JM (1985) Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res 13:1431–1442
Grunfeld C, Gulli R, Moser AH, Gavin LA, Feingold KR (1989) Effect of tumor necrosis factor administration in vivo on lipoprotein lipase activity in various tissues of the rat. J Lipid Res 30:579–585
Irikura T, Takagi K, Okada K, Yagasaki K (1985) Effect of KCD-232, a new hypolipidemic agent, on serum lipoprotein changes in hepatoma-bearing rats. Lipids 20:420–424
Kawakami M, Pekala PH, Lane MD, Cerami A (1982) Lipoprotein lipase suppression in 3T3–L1 cells by an endotoxin-induced mediator from exudate cells. Proc Natl Acad Sci USA 79:912–916
Kawamura M, Jensen DF, Wancewicz EV, Joy LL, Khoo JC, Steinberg D (1981) Hormone-sensitive lipase in differentiated 3T3–L1 cells and its activation by cyclic AMP-dependent protein kinase. Proc Natl Acad Sci USA 78:732–736
Kawasaki M, Yagasaki K, Miura Y, Funabiki R (1995) Reduction of hyperlipidemia in hepatoma-bearing rats by dietary fish oil. Lipids 30:431–436
Kawasaki M, Yagasaki K, Miura Y, Funabiki R (2004) Comparison of the changes in lipid metabolism between hepatoma-bearing and lipopolysaccharide-treated rats. Biosci Biotechnol Biochem 68:72–78
Kawasaki M, Miura Y, Funabiki R, Yagasaki K (2010) Comparison of the effects on lipid metabolism of dietary methionine and cystine between hepatoma-bearing and normal rats. Biosci Biotechnol Biochem 74:153–167
Muntoni S, Atzori L, Mereu R, Satta G, Macis MD, Congia M, Tedde A, Desogus A, Muntoni S (2009) Serum lipoproteins and cancer. Nutr Metab Cardiovasc Dis 19:218–225
Nakai T, Yamada S, Tamai T, Kobayashi T, Hayashi T, Takeda R (1979) The effects of streptozotocin diabetes on hepatic triglyceride lipase activity in the rat. Metabolism 28:30–40
Nilsson-Ehle P, Schotz MC (1976) A stable, radioactive substrate emulsion for assay of lipoprotein lipase. J Lipid Res 17:536–541
Nydegger UE, Butler RE (1972) Serum lipoprotein levels in patients with cancer. Cancer Res 32:1756–1760
Price SR, Olivecrona T, Pekala PH (1986) Regulation of lipoprotein lipase synthesis by recombinant tumor necrosis factor-the primary regulatory role of the hormone in 3T3–L1 adipocytes. Arch Biochem Biophys 251:738–746
Qadir MI, Malik SA (2008) Plasma lipid profile in gynecologic cancers. Eur J Gynaecol Oncol 29:158–161
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York, pp 7.53–7.55
Sugiyama K, Kushima Y, Muramatsu K (1984) Effects of methionine, cystine and taurine on plasma cholesterol level in rats fed a high cholesterol diet. Agric Biol Chem 48:2897–2899
Wion KL, Kirchgessner TG, Lusis AJ, Schotz MC, Lawn RM (1987) Human lipoprotein lipase complementary DNA sequence. Science 235:1638–1641
Yagasaki K, Okada K, Takagi K, Irikura T (1984) Effect of 4-(4′-chlorobenzyloxy) benzyl nicotinate (KCD-232) on cholesterol metabolism in rats fed an amino acid imbalance diet. Agric Biol Chem 48:1417–1423
Yagasaki K, Machida M, Funabiki R (1986) Effects of dietary methionine, cystine, and glycine on endogenous hypercholesterolemia in hepatoma-bearing rats. J Nutr Sci Vitaminol 32:643–651
Acknowledgments
We are indebted to Dr. Teruo Kawada, Kyoto University, for the gift of mouse 3T3-L1 cells, to Dr. Michael C. Schotz, University of California, Los Angeles, for presenting the LPL cDNA probe and to Dr. Toshio Watanabe, University of Tokyo, for providing the GAPDH cDNA probe.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kawasaki, M., Miura, Y. & Yagasaki, K. Effects of sulfur amino acids, l-methionine, l-cystine and l-cysteine on lipoprotein lipase and hormone-sensitive lipase in differentiated mouse 3T3-L1 adipocytes. Cytotechnology 62, 225–233 (2010). https://doi.org/10.1007/s10616-010-9282-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10616-010-9282-0