Role of Tumor Suppressor PTEN in Tumor Necrosis Factor α-Induced Inhibition of Insulin Signaling in Murine Skeletal Muscle C₂C₁₂ Cells

Abstract

In an attempt to clarify the role of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) in muscle insulin resistance, we investigated the effect of PTEN on phosphoinositide 3 (PI3)-kinase/Akt related insulin signaling pathway in skeletal muscle-like C₂C₁₂ cells damaged by tumor necrosis factor-alpha (TNFα). C₂C₁₂ cells cultured with TNFα (10 ng/ml) for 1 h displayed a marked decrease of insulin-stimulated 2-[¹⁴C]-deoxy-d-glucose (2-DG) uptake in parallel with an elevation of PTEN mRNA and protein levels. However, pretreatment of PTEN antisense oligonucleotide (AS) (1 μmol/l for 3 days) for specific inhibition of PTEN expression in C₂C₁₂ cells abolished the TNFα-induced changes in 2-DG uptake. Similar pretreatment with PTEN AS, but not with sense oligonucleotide (1 μmol/l for 3 days), eliminated the ability of TNFα to impair insulin-stimulated signals including p85 regulatory subunit of PI3-kinase expression and the degree of Akt serine phosphorylation as well as protein expression in glucose transporter subtype 4. Data taken from cultured C₂C₁₂ cells emphasize the negative regulatory of muscle PI3-kinase/Akt signaling pathways as the major substrate of PTEN but also support the concept that PTEN contributes to the development of insulin resistance in skeletal muscle.

Key words

C₂C₁₂ cells - insulin resistance - insulin signaling - PTEN - TNFα

References

• 1 Steck PA, Pershouse MA, Jasser SA, Yung WK, Lin H, Ligon AH, Langford LA, Baumgard ML, Hattier T, Davis T, Frye C, Hu R, Swedlund B, Teng DH, Tavtigian SV. Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers.  Nat Genet. 1997;  15 356-362
• 2 Li J, Yen C, Liaw D, Podsypanina K, Bose S, Wang SI, Puc J, Miliaresis C, Rodgers L, McCombie R, Bigner SH, Giovanella BC, Ittmann M, Tycko B, Hibshoosh H, Wigler MH, Parsons R. PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer.  Science. 1997;  275 1943-1947
• 3 Podsypanina K, Ellenson LH, Nemes A, Gu J, Tamura M, Yamada KM, Cordon-Cardo C, Catoretti G, Fisher PE, Parsons R. Mutation of Pten/Mmac1 in mice causes neoplasia in multiple organ systems.  Proc Natl Acad Sci USA. 1999;  96 1563-1568
• 4 Scheid MP, Woodgett JR. PKB/AKT: functional insights from genetic models. Nature Reviews.  Mol Biol Cell. 2001;  2 760-768
• 5 Cho H, Mu J, Kim JK, Thorvaldsen JL, Chu Q, Crenshaw EB 3rd, Kaestner KH, Bartolomei MS, Shulman GI, Birnbaum MJ. Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta).  Science. 2001;  292 1728-1731
• 6 Stambolic V, Suzuki A, de la Pompa JL, Brothers GM, Mirtsos C, Sasaki T, Ruland J, Penninger JM, Siderovski DP, Mak TW. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor Pten.  Cell. 1998;  95 29-39
• 7 Nakashima N, Sharma PM, Imamura T, Bookstein R, Olefsky JM. The tumor suppressor PTEN negatively regulates insulin signaling in 3T3-L1 adipocytes.  J Biol Chem. 2000;  275 12889-12895
• 8 Butler M, McKay RA, Popoff IJ, Gaarde WA, Witchell D, Murray SF, Dean NM, Bhanot S, Monia BP. Specific inhibition of PTEN expression reverses hyperglycemia in diabetic mice.  Diabetes. 2002;  51 1028-1034
• 9 Lo YT, Tsao CJ, Liu IM, Liou SS, Cheng JT. Increase of PTEN gene expression in insulin resistance.  Horm Metab Res. 2004;  36 662-666
• 10 DeFronzo RA, Gunnarsson R, Bjorkman O, Olsson M, Wahren J. Effects of insulin on peripheral and splanchnic glucose metabolism in noninsulin-dependent (type II) diabetes mellitus.  J Clin Invest. 1985;  76 149-155
• 11 Vaag A, Henriksen JE, Beck-Nielsen H. Decreased insulin activation of glycogen synthase in skeletal muscles in young nonobese Caucasian first-degree relatives of patients with non-insulin-dependent diabetes mellitus.  J Clin Invest. 1992;  89 782-788
• 12 Wijesekara N, Konrad D, Eweida M, Jefferies C, Liadis N, Giacca A, Crackower M, Suzuki A, Mak TW, Kahn CR, Klip A, Woo M. Muscle-specific Pten deletion protects against insulin resistance and diabetes.  Mol Cell Biol. 2005;  25 1135-1145
• 13 Yaffe D, Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle.  Nature. 1977;  270 725-727
• 14 del Aguila LF, Claffey KP, Kirwan JP. TNF-alpha impairs insulin signaling and insulin stimulation of glucose uptake in C2C12 muscle cells.  Am J Physiol. 1999;  276 E849-E855
• 15 Ko WC, Liu TP, Cheng JT, Tzeng TF, Liu IM. Effect of opioid mu-receptors activation on insulin signals damaged by tumor necrosis factor alpha in myoblast C2C12 cells.  Neurosci Lett. 2006;  397 274-278
• 16 Hotamisligil GS, Arner P, Caro JF, Atkinson RL, Spiegelman BM. Increased adipose tissue expression of tumor necrosis factor-α in human obesity and insulin resistance.  J Clin Invest. 1995;  95 2409-2415
• 17 Lang CH, Dobrescu C, Bagby GJ. Tumor necrosis factor impairs insulin action on peripheral glucose disposal and hepatic glucose output.  Endocrinology. 1992;  130 43-52
• 18 Dias N, Stein CA. Antisense oligonucleotides: basic concepts and mechanisms.  Mol Cancer Ther. 2002;  1 347-355
• 19 Moe-Behrens GH, Klinger FG, Eskild W, Grotmol T, Haugen TB, De Felici M. Akt/PTEN signaling mediates estrogen-dependent proliferation of primordial germ cells in vitro.  J Mol Endocrinol. 2003;  17 2630-2638
• 20 Ballotti R, Kowalski A, White MF, Le Marchand-Brustel Y, Van Obberghen E. Insulin stimulates tyrosine phosphorylation of its receptor beta-subunit in intact rat hepatocytes.  Biochem J. 1987;  241 99-104
• 21 Yeh KT, Chang JG, Chen YJ, Chen ST, Yu SY, Shih MC, Perng LI, Wang JC, Tsai M, Chang CP. Mutation analysis of the putative tumor suppressor gene PTEN/MMAC1 in hepatocellular carcinoma.  Cancer Invest. 2000;  18 123-129
• 22 Kruszynska YT, Olefsky JM. Cellular and molecular mechanisms of non-insulin dependent diabetes mellitus.  J Investig Med. 1996;  44 413-428
• 23 Vasudevan KM, Gurumurthy S, Rangnekar VM. Suppression of PTEN expression by NF-κB prevents apoptosis.  Mol Cell Biol. 2004;  24 1007-1021
• 24 Shulman GI. Cellular mechanisms of insulin resistance.  J Clin Invest. 2000;  106 171-176
• 25 Carvalho E, Rondinone C, Smith U. Insulin resistance in fat cells from obese Zucker rats-evidence for an impaired activation and translocation of protein kinase B and glucose transporter 4.  Mol Cell Biol. 2000;  206 7-16
• 26 Kurokawa K, Oka Y. Insulin resistance and glucose transporter.  Nippon Rinsho. 2000;  58 310-314

Correspondence