Abstract
Aim:
To study the effect of peroxisome proliferator-actived receptor γ (PPARγ) ligands on cell proliferation and apoptosis in human renal carcinoma cell lines.
Methods:
The expression of PPARγ was investigated by reverse transcriptase polymerase chain reaction (RT-PCR), Western blot and immunohistochemistry. The effect of thiazolidinedione (TZD) PPARγ ligands on growth of renal cell carcinoma (RCC) cells was measured by MTT assay and flow cytometric analysis. Cell death ELISA, Hoechst 33342 fluorescent staining and DNA ladder assay were used to observe the effects of PPARγ ligands on apoptosis. Regulatory proteins of cell cycle and apoptosis were detected by Western blot analysis.
Results:
PPARγ was expressed at much higher levels in renal tumors than in the normal kidney (2.16±0.85 vs 0.90±0.73; P<0.01). TZD PPARγ ligands inhibited RCC cell growth in a dose-dependent manner with IC50 values of 7.08 μmol/L and 11.32 μmol/L for pioglitazone, and 5.71 μmol/L and 8.38 μmol/L for troglitazone in 786-O and A498 cells, respectively. Cell cycle analysis showed a G0/G1 arrest in human RCC cells following 24-h exposure to TZD. Analysis of cell cycle regulatory proteins revealed that TZD decreased the protein levels of proliferating cell nuclear antigen, pRb, cyclin D1, and Cdk4 but increased the levels of p21 and p27 in a time-dependent manner. Furthermore, high doses of TZD induced massive apoptosis in renal cancer cells, with increased Bax expression and decreased Bcl-2 expression.
Conclusion:
TZD PPARγ ligands showed potent inhibitory effect on proliferation, and could induce apoptosis in RCC cells. These results suggest that ligands for PPARγ have potential antitumor effects on renal carcinoma cells.
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References
Issemann I, Green S . Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. Nature 1990; 347: 645–50.
Schoonjans K, Martin G, Staels B, Auwerx J . Peroxisome proliferator-activated receptors, orphans with ligands and functions. Curr Opin Lipidol 1997; 8: 159–66.
Kliewer SA, Umesono K, Noonan DJ, Heyman RA, Evans RM . Convergence of 9–cis retinoic acid and peroxisome proliferator signaling pathways through heterodimer formation of their receptors. Nature 1992; 358: 771–4.
Kliewer SA, Lehmann JM, Milburn MV, Willson TM . The PPARs and PXRs: nuclear xenobiotic receptors that define novel hormone signaling pathways. Recent Prog Horm Res 1999; 54: 345–67.
Palmer CN, Hsu MH, Griffin HJ, Johnson EF . Novel sequence determinants in peroxisome proliferator signaling. J Biol Chem 1995; 270: 16114–21.
Camp HS, Ren D, Leff T . Adipogenesis and fat-cell function in obesity and diabetes. Trends Mol Med 2002; 8: 442–7.
Rangwala SM, Lazar MA . Peroxisome proliferator-activated receptor gamma in diabetes and metabolism. Trends Pharmacol Sci 2004; 25: 331–6.
Fauconnet S, Lascombe I, Chabannes E, Adessi GL, Desvergne B, Wahli W, et al. Differential regulation of vascular endothelial growth factor expression by peroxisome proliferator-activated receptors in bladder cancer cells. J Biol Chem 2002; 277: 23534–43.
Sarraf P, Mueller E, Jones D, King FJ, DeAngelo DJ, Partridge JB, et al. Differentiation and reversal of malignant changes in colon cancer through PPARgamma. Nat Med 1998; 4: 1046–52.
Elstner E, Muller C, Koshizuka K, Williamson EA, Park D, Asou H, et al. Ligands for peroxisome proliferator-activated receptor-gamma and retinoic acid receptor inhibit growth and induce apoptosis of human breast cancer cells in vitro and in BNX mice. Proc Natl Acad Sci USA 1998; 95: 8806–11.
Heaney AP, Fernando M, Melmed S . PPAR-gamma receptor ligands: novel therapy for pituitary adenomas. J Clin Invest 2003; 111: 1381–8.
Takahashi N, Okumura T, Motomura W, Fujimoto Y, Kawabata I, Kohgo Y . Activation of PPARgamma inhibits cell growth and induces apoptosis in human gastric cancer cells. FEBS Lett 1999; 455: 135–9.
Guan YF, Zhang YH, Breyer RM, Davis L, Breyer MD . Expression of peroxisome proliferator-activated receptor gamma (PPARgamma) in human transitional bladder cancer and its role in inducing cell death. Neoplasia 1999; 1: 330–9.
Demetri GD, Fletcher CD, Mueller E, Sarraf P, Naujoks R, Campbell N, et al. Induction of solid tumor differentiation by the peroxisome proliferator-activated receptor-γ ligand troglitazone in patients with liposarcoma. Proc Natl Acad Sci USA 1999; 96: 3951–6.
Mueller E, Smith M, Sarraf P, Kroll T, Aiyer A, Kaufman DS, et al. Effects of ligand activation of peroxisome proliferator-activated receptor gamma in human prostate cancer. Proc Natl Acad Sci USA 2000; 97; 10990–5.
Golimbu M, Joshi P, Sperber A, Tessler A, Al-Askari S, Morales P . Renal cell carcinoma: survival and prognostic factors. Urology 1986; 27: 291–301.
Flanigan RC, Campbell SC, Clark JI, Picken MM . Metastatic renal cell carcinoma. Curr Treat Options Oncol 2003; 4: 385–90.
Guan Y, Zhang Y, Schneider A, Davis L, Breyer RM, Breyer MD . Peroxisome proliferator-activated receptor-gamma activity is associated with renal microvasculature. Am J Physiol Renal Physiol 2001; 281: 1036–46.
Motomura W, Okumura T, Takahashi N, Obara T, Kohgo Y . Activation of peroxisome proliferator-activated receptor gamma by troglitazone inhibits cell growth through the increase of p27KiP1 in human pancreatic carcinoma cells. Cancer Res 2000; 60: 5558–64.
Hashimoto Y, Shimada Y, Itami A, Ito T, Kawamura J, Kawabe A, et al. Growth inhibition through activation of peroxisome proliferator-activated receptor gamma in human oesophageal squamous cell carcinoma. Eur J Cancer 2003; 39: 2239–46.
Inoue K, Kawahito Y, Tsubouchi Y, Kohno M, Yoshimura R, Yoshikawa T, et al. Expression of peroxisome proliferator-activated receptor gamma in renal cell carcinoma and growth inhibition by its agonists. Biochem Biophys Res Commun 2001; 287: 727–32.
Heaney AP, Fernando M, Yong WH, Melmed S . Functional PPAR-gamma receptor is a novel therapeutic target for ACTH-secreting pituitary adenomas. Nat Med 2002; 8: 1281–7.
Mossner R, Schulz U, Kruger U, Middel P, Schinner S, Fuzesi L, et al. Agonists of peroxisome proliferator-activated receptor gamma inhibit cell growth in malignant melanoma. J Invest Dermatol 2002; 119: 576–82.
Brockman JA, Gupta RA, Dubois RN . Activation of PPARgamma leads to inhibition of anchorage-independent growth of human colorectal cancer cells. Gastroenterology 1998; 115: 1049–55.
Luo Y, Hurwitz J, Massague J . Cell-cycle inhibition by independent CDK and PCNA binding domains in p21Cip1. Nature 1995; 375: 159–61.
Pines J . Four-dimensional control of the cell cycle. Nat Cell Biol 1999; 1: 73–9.
Bartek J, Bartkova J, Lukas J . The retinoblastoma protein pathway in cell cycle control and cancer. Exp Cell Res 1997; 237: 1–6.
Sherr CJ . Mammalian G1 cyclins. Cell 1993; 73: 1059–65.
Aprelikova O, Xiong Y, Liu ET . Both p16 and p21 families of cyclin-dependent kinase (CDK) inhibitors block the phosphorylation of cyclin-dependent kinases by the CDK-activating kinase. J Biol Chem 1995; 270: 18195–7.
Shi C, Yu L, Yang F, Yan J, Zeng H . A novel organoselenium compound induces cell cycle arrest and apoptosis in prostate cancer cell lines. Biochem Biophys Res Commun 2003; 309: 578–83.
Zhang CL, Wu LJ, Tashiro S, Onodera S, Ikejima T . Oridonin induces apoptosis of HeLa cells via altering expression of Bcl-2/ Bax and activating caspase-3/ICAD pathway. Acta Pharmacol Sin 2004; 25: 691–8.
Filippova M, Parkhurst L, Duerksen-Hughes PJ . The human papillomavirus 16 E6 protein binds to Fas-associated death domain and protects cells from Fas-triggered apoptosis. J Biol Chem 2004; 279: 25729–44.
Orrenius S . Mitochondrial regulation of apoptotic cell death. Toxicol Lett 2004; 149: 19–23.
Li XH, Li JJ, Zhang HW, Sun P, Zhang YL, Cai SH, et al. Nimesulide inhibits tumor growth in mice implanted hepatoma: overexpression of Bax over Bcl-2.
Salomons GS, Brady HJ, Verwijs-Janssen M, Van Den Berg JD, Hart AA, Van Den Berg H, et al. The Bax alpha:Bcl-2 ratio modulates the response to dexamethasone in leukaemic cells and is highly variable in childhood acute leukaemia. Int J Cancer 1997; 71: 959–65.
Chawla A, Barak Y, Nagy L, Liao D, Tontonoz P, Evans RM . PPAR-gamma dependent and independent effects on macrophage-gene expression in lipid metabolism and inflammation. Nat Med 2001; 7: 48–52.
Davies GF, Khandelwal RL, Wu L, Juurlink BH, Roesler WJ . Inhibition of phosphoenolpyruvate carboxykinase (PEPCK) gene expression by troglitazone: a peroxisome proliferator-activated receptor-gamma (PPARgamma)-independent, antioxidant-related mechanism. Biochem Pharmacol 2001; 62: 1071–9.
Wang M, Wise SC, Leff T, Su TZ . Troglitazone, an antidiabetic agent, inhibits cholesterol biosynthesis through a mechanism independent of peroxisome proliferator-activated receptor-gamma. Diabetes 1999; 48: 254–60.
Wang YL, Frauwirth KA, Rangwala SM, Lazar MA, Thompson CB . Thiazolidinedione activation of peroxisome proliferator-activated receptor gamma can enhance mitochondrial potential and promote cell survival. J Biol Chem 2002; 277: 31781–8.
Yu C, Chen L, Luo H, Chen J, Cheng F, Gui C, et al. Binding analyses between human PPARgamma-LBD and ligands. Eur J Biochem 2004; 271: 386–97.
Bae MA, Rhee H, Song BJ . Troglitazone but not rosiglitazone induces G1 cell cycle arrest and apoptosis in human and rat hepatoma cell lines. Toxicol Lett 2003; 139: 67–75.
Bova MP, Tam D, McMahon G, Mattson MN . Troglitazone induces a rapid drop of mitochondrial membrane potential in liver HepG2 cells. Toxicol Lett 2005; 155: 41–50.
Tsuchiya T, Shimizu H, Shimomura K, Mori M . Troglitazone inhibits isolated cell proliferation, and induces apoptosis in isolated rat mesangial cells. Am J Nephrol 2003; 23: 222–8.
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Project partly supported by National Natural Science Foundation of China (No 30250002 to Yin-lu GUO and No 30271521, No 30340084 and No G2000056908 to You-fei GUAN) and a grant from Peking University “211” Program (to You-fei GUAN).
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Yang, Fg., Zhang, Zw., Xin, Dq. et al. Peroxisome proliferator-activated receptor γ ligands induce cell cycle arrest and apoptosis in human renal carcinoma cell lines. Acta Pharmacol Sin 26, 753–761 (2005). https://doi.org/10.1111/j.1745-7254.2005.00753.x
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DOI: https://doi.org/10.1111/j.1745-7254.2005.00753.x
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