罗格列酮对体外培养人肝癌SMMC7721细胞生长的影响
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摘要
目的 探讨过氧化物酶体增殖因子活化受体γ(Peroxisome Proliferator-actived Recepter PPARγ)配体罗格列酮(Rosiglitazone，RSG)体外抑制人肝癌SMMC7721细胞系细胞生长作用及初步探讨其作用机制。
方法 体外培养人肝癌SMMC7721细胞，以全反式维甲酸(All-trans retinoic，ATRA)为阳性对照药，MTT比色试验测定药物对细胞生长的影响，流式细胞术(Flow cytometry，FCM)检测细胞周期和细胞凋亡，分光光度法测定γ-GT变化，ELISA检测AFP分泌量，免疫细胞化学检测药物处理前后PPARγ，CyclinD_1蛋白表达的改变。
结果 不同浓度的RSG(12.5、25、50、100μmol／L)和10μmol／L的ATRA处理细胞后抑制肝癌SMMC7721细胞生长，RSG抑制肝癌SMMC7721细胞生长呈时间依赖性和浓度依赖性；γ-GT量明显下降，AFP分泌显著降低；G1期细胞数目增加，S期和G_2数目减少，而凋亡率无明显改变；细胞的PPARγ蛋白核内表达上调，CyclinD_1蛋白表达下调。
结论 过氧化物酶体增殖因子活化受体γ(PPARγ)配体罗格列酮(Rosiglitazone，RSG)具有抑制体外培养肝癌SMMC7721细胞生长作用；罗格列酮对SMMC7721细胞生长抑制作用与其阻滞细胞周期和诱导细胞分化相关；PPARγ蛋白核内表达上调、CyclinD_1蛋白表达下调可能介导了RSG抑制细胞生长。
Objective To investigate the effect of Rosiglitazone , the ligand of
Peroxisome Proliferator-activated Receptor gamma, on cell proliferation inhibition of hepatocarcinoma cells in vitro and its possible mechanism.
Methods Human hepatocarcinoma SMMC7721 cell line was cultured in
vitro, Cultured SMMC7721 cells were designed into 12.5 μ mol/L RSG group, 25 μ mol/L RSG group, 50 μ mol/L RSG group, 100 μ mol/L RSG group and the control group, 10 μ mol/L ATRA group. Proliferation inhibition effect was tested by MTT assay; Flow cytometry was used to detect cell cycle and apoptosis rate; The activity of γ -GT was measured with γ -GT kit and the secretive AFP was measured with ELISA ; Expression of PPARγ、 CyclinD_1 protein in hepatocarcinoma cells between untreated and treated with or without Rosiglitazone were observed by immunohistochemistry.
Results Rosiglitazone at various concentrations of 12.5、 25、 50、 100
μmol/L and All-trans retinoic acid at concentration 10 μmol·L~(-1) could inhibit the proliferation of SMMC7721 cells, RSG inhibit the proliferation in a dose-dependent and time-dependent manner; Cell micrographys tended to be normal after treatment; the specific activities of γ-GT and the secretive amount of AFP was reduced; Flow cytometry analysis reavealed a cell cycle arrest at G1 phase ,but apoptosis rate was not changed remarkably; nuclear traslocation and up-regulation of PPARγ protein expression and the down -regulation of CyclinD protein expression were observed .
Conclusion The ligand of PPARγ, Rosiglitazone induced significantly growth inhibition of human hepatocarcinoma SMMC7721 cells line in vitro which
方法 体外培养人肝癌SMMC7721细胞，以全反式维甲酸(All-trans retinoic，ATRA)为阳性对照药，MTT比色试验测定药物对细胞生长的影响，流式细胞术(Flow cytometry，FCM)检测细胞周期和细胞凋亡，分光光度法测定γ-GT变化，ELISA检测AFP分泌量，免疫细胞化学检测药物处理前后PPARγ，CyclinD_1蛋白表达的改变。
结果 不同浓度的RSG(12.5、25、50、100μmol／L)和10μmol／L的ATRA处理细胞后抑制肝癌SMMC7721细胞生长，RSG抑制肝癌SMMC7721细胞生长呈时间依赖性和浓度依赖性；γ-GT量明显下降，AFP分泌显著降低；G1期细胞数目增加，S期和G_2数目减少，而凋亡率无明显改变；细胞的PPARγ蛋白核内表达上调，CyclinD_1蛋白表达下调。
结论 过氧化物酶体增殖因子活化受体γ(PPARγ)配体罗格列酮(Rosiglitazone，RSG)具有抑制体外培养肝癌SMMC7721细胞生长作用；罗格列酮对SMMC7721细胞生长抑制作用与其阻滞细胞周期和诱导细胞分化相关；PPARγ蛋白核内表达上调、CyclinD_1蛋白表达下调可能介导了RSG抑制细胞生长。
Objective To investigate the effect of Rosiglitazone , the ligand of
Peroxisome Proliferator-activated Receptor gamma, on cell proliferation inhibition of hepatocarcinoma cells in vitro and its possible mechanism.
Methods Human hepatocarcinoma SMMC7721 cell line was cultured in
vitro, Cultured SMMC7721 cells were designed into 12.5 μ mol/L RSG group, 25 μ mol/L RSG group, 50 μ mol/L RSG group, 100 μ mol/L RSG group and the control group, 10 μ mol/L ATRA group. Proliferation inhibition effect was tested by MTT assay; Flow cytometry was used to detect cell cycle and apoptosis rate; The activity of γ -GT was measured with γ -GT kit and the secretive AFP was measured with ELISA ; Expression of PPARγ、 CyclinD_1 protein in hepatocarcinoma cells between untreated and treated with or without Rosiglitazone were observed by immunohistochemistry.
Results Rosiglitazone at various concentrations of 12.5、 25、 50、 100
μmol/L and All-trans retinoic acid at concentration 10 μmol·L~(-1) could inhibit the proliferation of SMMC7721 cells, RSG inhibit the proliferation in a dose-dependent and time-dependent manner; Cell micrographys tended to be normal after treatment; the specific activities of γ-GT and the secretive amount of AFP was reduced; Flow cytometry analysis reavealed a cell cycle arrest at G1 phase ,but apoptosis rate was not changed remarkably; nuclear traslocation and up-regulation of PPARγ protein expression and the down -regulation of CyclinD protein expression were observed .
Conclusion The ligand of PPARγ, Rosiglitazone induced significantly growth inhibition of human hepatocarcinoma SMMC7721 cells line in vitro which
引文
1. Esdar C, Milasta S, Maelicke A, et al. Differentiation-associatal apoptosis of neural stem cells is effected by Bcl-2 overexpression: impacton cell lineage determination[J]. Eur J Cell Biol, 2001; 80(8): 539-553.
2. ZhaoX, Demary K, WongL, et al. Retinoic acid receptor-independent mechanism of apoptosis of melanoma cells by the retinoid CD437(AHPN) [J]. Cell Death Differ,2001;8(9):878-886.
3. Kliewer SA, Umesono K,Noonan DJ, et al. Convergence of 9-cis retinoic acid and peroxisome proliferator signaling pathways through heterodimer formation of their receptors Nature. 1992 Aug 27;358(6389):771-774.
4. Marcus SL,Capone JP,Rachubinski RA.Identification of COUP-TFII as a Peroxisome proliferator response element binding factor using genetic seletion I yeast: COUP-TFII activates transcription in yeast but antagonizes PPAR signaling in mammalian cells. Mol Cell Endocrinol 1996, 120: 31- 39.
5. Rao MS, Reddy JK. Peroxisomal beta-oxidation and steatohepatitis. Semin Liver Dis. 2001; 21(1):43-55.
6. Daynes RA,Jones DC.Emerging roles of PPARs in inflammation and immunity.Nat Rev Immunol.2002 Oct;2(10):748-759.
7. Miyahara T,Schrum L,Rippe R,et al. Peroxisome proliferator-activated receptors and hepatic stellate cell activation.J Biol Chem.2000 Nov 17;275(46):35715-35722.
8. Scheen AJ, Paquot N. PPAR-gamma receptors, new therapeutic target in metabolic and cardiovascular diseasesRev Med Liege. 2005 Feb; 60(2):89-95.
9. Martens FM, Visseren FL, Lemay J, et al. Metabolic and additional vascular effects of thiazolidinediones. Drugs. 2002; 62(10): 1463-1480.
10. Shouwei Han, Neil Sidell, Paul B, et al. Up-regulation of p21 Gene Expression by Peroxisome Proliferator-Activated Receptorγ in Human Lung Carcinoma Cells. Clin Cancer Res. 2004, Mar 15; 10(6):1911-1919.
11. Reed JC. Apoptosis mechanisms: implications for cancer drug discovery.??Oncology (Huntingt). 2004 Nov; 18(13 Suppl 10):11-20.
12. Yoshimura R, Matsuyama M, Segawa Y et al. Expression of peroxisome proliferator-activated receptors (PPARs) in human urinary bladder carcinoma and growth inhibition by its agonists. Int J Cancer. 2003 May 1; 104(5): 5 97-602.
13. Guan YF, Zhang YH, Breyer RM et al. Expression of peroxisome proliferator-activated receptor gamma (PPARgamma) in human transitional bladder cancer and its role in inducing cell death. Neoplasia. 1999 Oct;1(4):330-339.
14. Heaney AP, Fernando M, Melmed S. PPAR-gamma receptor ligands: novel therapy for pituitary adenomas. J Clin Invest. 2003 May; 111(9): 1381-8.
15. Keshamouni VG, Arenberg DA, Reddy RC, et al. PPAR-gamma activation inhibits angiogenesis by blocking ELR+CXC chemokine production in non-small cell lung cancer. Neoplasia. 2005 Mar;7(3):294-301
16. Lee KS, Park SJ, Hwang PH, et al. PPAR-gamma modulates allergic inflammation through up-regulation of PTEN. FASEBJ.2005 Jun; 19(8): 1033-5.
17. Konturek PC, Kania J, Konturek JW, et al. H. pylori infection, atrophic gastritis, cytokines, gastrin, COX-2, PPAR gamma and impaired apoptosis in gastric carcinogenesis. Med Sci Monk. 2003 Jul; 9(7):SR53-66.
18. Guan Y. Targeting peroxisome proliferator-activated receptors (PPARs) in kidney and urologic disease. Minerva Urol Nefrol. 2002 Jun; 54(2):65-79.
19. Wijnhoven BP, Lindstedt EW, Abbou M et alJ Dinjenens, WN.Molecular genetic analysis of the von Hippel—Lindau and human Peroxisome Proliferator-activated receptor gamma tumor-suppressor genes in adenocarcinomas of the gastroesophageal junction. Int J Cancer, 2001; 94(6): 891—895
20. Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of peroxisome proliferator-activated receptor gamma in non-small cell lung Cancer.Cancer Res, 2000; 60(4): 1129-1138.
21. HeTC,ChanTA,VogelsteinB, et al. PPARγis an APC-Regulated Target of??Nonsteroidal Anti-InflammatoryDrugs. Cell, 1999: 336～345.
22. Smith SA. Peroxisome proliferator-activated receptors and the regulation of mammalian lipid metabolism. Biochem Soc Trans. 2002 Nov; 30(Pt 6): 1086-1090.
23. Altiok S, Xu M, Spiegelman BM. PPARgamma induces cell cycle withdrawal: inhibition of E2F/DP DNA-binding activity via down-regulation of PP2A. Genes Dev. 1997Aug 1; 11(15): 1987-1998.
24. Tsubouchi Y, Sano H, Kawahito Y et al. Inhibition of human lung cancer cell growth by the peroxisome proliferator-activated receptor-gamma agonists through induction of apoptosis. Biochem Biophys Res Commun. 2000 Apr 13; 270(2):400-405.
25．周亭芳，庄英帜，曹建国．罗格列酮增强顺铂抑制人肺腺癌细胞增殖作用[J]中国药理学通报，2005，21(1)：88-91．
26. Sherr CJ. The Pezzoller Lecture: cancer cell cycle revisited. Cancer Res, 200; 60: 3689.
27. Koga H, Sakisaka S, Harada M, et al. Involvement of p21(WAF1/Cip1), p27(Kip1), and p18(INK4c) in troglitazoneinduced cell-cycle arrest in human hepatoma cell lines. Hepatology 2001; 33: 1087-1097.
28. Itami A, Watanabe G, Shimada Y, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth of pancreatic cancers both in vitro and in vivo. Int J Cancer 2001; 94: 370-376.
29. Motomura W, Okumura T, Takahashi N, et al. 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: 5558-5564.
30. Yin F, Wakino S, Liu Z, et al. Troglitazone inhibits growth of MCF-7 breast carcinoma cells by targeting G1 cell-cycle regulators. Bioehem Biophys Res Commun 2001; 286: 916-922.
31. Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of peroxisome proliferator-activated receptor gamma in nonsmall-cell lung cancer.??Cancer Res 2000; 60:1129-1138.
32. Morrison RF, Farmer SR. Role of PPARgamma in regulating a cascade expression of cyclin-dependent kinase inhibitors, p18(INK4c) and p21(Waf1/Cip1), during adipogenesis. J Biol Chem 1999; 274:17088-17097.
33. Elnemr A, Ohta T, Iwata K, et al. PPARgamma ligand (thiazolidinedione) induces growth arrest and differentiation markers of human pancreatic cancer cells. Int J Oncol 2000; 17:1157-1164.
34. Itami A, Watanabe G, Shimada Y, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth of pancreatic cancers both in vitro and in vivo. Int J Cancer 2001; 94: 370-376.
35. Tien HF, Wang CH. CD7 positive hematopoietic progenitors and acute myeloid leukemia and other minimally differentiated leukemia. Leuk Lymphoma. 1998 Sep; 31(1-2):93-98.
36. samura RY, Egashira N, Miyai Set al.Molecular pathology of the pituitary. Development and functional differentiation of pituitary adenomas. Front Horm Res. 2004; 2:20-33.
37. Asou H, Verbeek W, Williamson E, et al. Growth inhibition of myeloid leukemia cells by troglitazone, a ligand for peroxisome proliferator activated receptor gamma, and retinoids. Int J Oncol 1999; 15:1027-1031.
38. Ray DM, Bernstein SH, Phipps RP. Human multiple myeloma cells express peroxisome proliferator-activated receptor gamma and undergo apoptosis upon exposure to PPARgamma ligands. Clin Immunol. 2004 Nov; 113(2):203-213.
39．董育玮，王兴鹏，吴恺 et al．过氧化物酶增殖物活化受体γ在胰腺癌生长中的调节作用．中华内科杂志，2003，42(7)：479-482．
40. Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of peroxisome proliferator-activated receptor gamma in non-small cell lung Cancer. Cancer Res, 2000; 60(4): 1129-1138.
41．曾益新编(主编)肿瘤学，第三版．北京：人民卫生出版社，2003：201．
42. Pitot HC, Glauert HP, Hanigan M. The significance of selected biochemical markers in the characterization of putative initiated cell populations in rodent liver.??Cancer Lett. 1985 Oct; 29(1):1-14.
43．秦锁富，查锡良，陈惠黎．亚硒酸钠对人肝癌细胞株甲胎蛋白纤维连接蛋白及细胞膜糖链的影响[J]．肿瘤，1991，11(3)：133．
44. Cox PJ, Ryan DA, Hollis FJ, et al. Absorption, disposition, and metabolism of rosiglitazone, a potent thiazolidinedione insulin sensitizer, in humans. Drug Metab Dispos, 2000; 28(7): 772-780.1. Michalik L, Wahli W. Peroxisome proliferator-activated receptors:three isotypes for a multitude of functions. [J]. Curr Opin Biotechnol. 1999 Dec; 10(6):564-570.
2. Issemann I, Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. [J]. Nature. 1990 Oct 18; 347(6294):645-650.
3 Farrow B, Evers BM. Activation of PPARgamma increases PTEN expression in??pancreatic cancer cells. (J) . Biochem Biophys Res Commun. 2003 Jan 31; 301(1):50-53.
4 Berger J, Moller DE. The mechanisms of action of PPARs. [J]. Annu Rev Med. 2002; 53: 409-435.
5 Subbaramaiah K, Lin DT, Hart JC,et al. Peroxisomeproliferator-actived receptor gamma ligands suppress the transcriptional activation of cyclooxygenase-2. Evidence for involvement of activator protein-1 and CREB-binding protein/p300.J Biol Chem. 2001 Apr 13; 276(15): 12440-12778. Epub 2001 Jan 23.
6 Zander T, Kraus JA, Grommes C, et al. Induction of apoptosis in human and rat glioma by agonists of the nuclear receptor PPARgamma. J Neurochem 2002; 81: 1052-1060.
7 Chattopadhyay N, Singh DP, Heese O, et al. Expression of peroxisome proliferator-activated receptors (PPARS) in human astrocytic cells: PPARgamma agonists as inducers of apoptosis.J Neurosci Res 2000; 61: 61-74.
8 Yoshida K, Tanabe K, Fujii D, et al.Induction mechanism of apoptosis by troglitazone through peroxisome proliferator-activated receptor-gamma in gastric carcinoma cells. [J]. Anticancer Res. 2003 Jan-Feb; 23(1A):267-273.
9 Shimada T, Kojima K, Yoshiura K, et al. Characteristics of the peroxisome proliferator-activated receptor gamma (PPARgamma)ligand-induced apoptosis in colon cancer cells. Gut 2002; 50: 658-664.
10 Eibl G, Wente MN, Reber HA, Hines OJ. Peroxisome proliferatoractivated receptor gamma induces pancreatic cancer cell apoptosis. Biochem Biophys Res Commun 2001; 287: 522-529.
11 Toyoda M, Takagi H, Horiguchi N, et al. A ligand for peroxisome proliferator-activated receptor gamma inhibits cell growth and induces apoptosis in human liver cancer cells. Gut 2002; 50: 563-567.
12 Ohta K, Endo T, Haraguchi K, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth and induce apoptosis of human papillary thyroid carcinoma cells. J Clin Endocrinol Metab 2001; 86: 2170-2177.
13 Tontonoz P, Hu E, Spiegelman BM. Stimulation of adipogenesis in fibroblasts by??PPAR gamma 2, a lipid-activated transcription factor. Cell 1994; 79: 1147-1156.
14 Okuno A, Tamemoto H, Tobe K, et al. Troglitazone increases the number of small adipocytes without the change of white adipose tissue mass in obese Zucker rats. J Clin Invest 1998; 101: 1354-1361.
15 Tontonoz P, Singer S, Forman BM, et al. Terminal differentiation of human liposarcoma cells induced by ligands for peroxisome proliferator-activated receptor gamma and the retinoid X receptor.Proc Natl Acad Sci USA 1997; 94: 237-241.
16 Elnemr A, Ohta T, Iwata K, et al. PPARgamma ligand (thiazolidinedione) induces growth arrest and differentiation markers of human pancreatic cancer cells. Int J Oncol 2000; 17: 1157-1164.
17 Altiok S, Xu M, Spiegelman BM. PPARgamma induces cell cycle withdrawal: inhibition of E2F/DP DNA-binding activity via down-regulation of PP2A. Genes Dev. 1997 Aug 1; 11(15): 1987-1998.
18 Morrison RF, Farmer SR. Role of PPARgamma in regulating a cascade expression of cyclin-dependent kinase inhibitors, p18(INK4c) and p21(Waf1/Cip1), during adipogenesis. J Biol Chem 1999; 274: 17088-97.
19 Koga H, Sakisaka S, Harada M, et al. Involvement of p21(WAF1/Cip1), p27(Kip1), and p18(INK4c) in troglitazoneinduced cell-cycle arrest in human hepatoma cell lines. Hepatology2001; 33: 1087-1097.
20 Itami A, Watanabe G, Shimada Y, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth of pancreatic cancers both in vitro and in vivo. Int J Cancer 2001; 94: 370-376.
21 Motomura W, Okumura T, Takahashi N, et al. 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: 5558-5564.
22 Yin F, Wakino S, Liu Z, et al. Troglitazone inhibits growth of MCF-7 breast carcinoma cells by targeting G1 cell-cycle regulators. Biochem Biophys Res Commun 2001; 286: 916-922.
23 Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of??peroxisome proliferator-activated receptor gamma in nonsmall-cell lung cancer. Cancer Res 2000; 60: 1129-1138.
24 XinX, Yang S, Kowalski J, Gerritsen ME. Peroxisome proliferatoractivated receptor gamma ligands are potent inhibitors of angiogenesis in vitro and in vivo. J Biol Chem 1999; 274: 9116-9121.
25 Panigrahy D, Singer S, Shen LQ, et al. PPARgamma ligands inhibit primary-tumor growth and metastasis by inhibiting angiogenesis. J Clin Invest 2002; 110: 923-932.
26 Panigrahy D, Dulak J, Piatkowska E, et al. Ligands of peroxisome proliferator-activated receptor-gamma increase the generation of vascular endothelial growth factor in vascular smooth-muscle cells and in macrophages. Acta Biochim Pol 2000; 47: 1147-1157.
27 Benjamin L, Golijanin D, Itin A et al. Selective ablation of immature blood vessels in established tumours follows vascular endothelial growth factor withdrawal. J Clin Invest 1999; 103: 159-165.
28 Freed MI, Allen A, Jorkasky DK, DiCicco RA. Systemic exposure to rosiglitazone is unaltered by food. Eur J Clin Pharmacol 1999;55: 53-56.
29 Sierra-Honigmann MR, Nath AK, Murakami C et al. Biological action of leptin as an angiogenic factor. Science 1998; 281: 1683- 1686.
30 Goetze S, Bungenstock A, Czupalla C, et al. Leptin induces endothelial-cell migration through Akt, which is inhibited by PPARgamma-ligands. Hypertension 2002; 40: 748-754.
31 Cui Y., Miyoshi K., Claudio E., Siebenlist U. K., Gonzalez F. J., Flaws J., Wagner K-U., Hennighausen L. Loss of the peroxisome proliferation-activated receptor gamma (PPAR(?)) does not affect mammary development and propensity for tumor formation but leads to reduce fertility. J. Biol. Chem., 277: 17830-17835, 2002.
32 Takayuki Ikezoe, Carl W, et al. Mutational Analysis of the Peroxisome Proliferator-activated Receptor (?) Gene in Human Malignancies. Cancer Research 61, 2001, July 1, 5307-5310.
33 Palakurthi S. S., Aktas H., Grubissich L. M., Mortensen R. M., Halperin J. A. ??Anticancer effects of thiazolidinediones are independent of peroxisome proliferator-activated receptor γand mediated by inhibition of translation initiation. Cancer Res., 61: 6213-6218, 2001.
34 Rossi A., Kapahi P., Natoli G, Takahashi T., Chen Y., Karin M., Santoro M. G. Anti-inflammatory cyclopentenone prostaglandins are direct inhibitors of INB kinase. Nature (Lond.), 403: 103-106, 2000.
35 Palakurthi S. S., Aktas H., Grubissich L. M., Mortensen R. M., Halperin J. A. Anticancer effects of thiazolidinediones are independent of peroxisome proliferator-activated receptor γand mediated by inhibition of translation initiation. Cancer Res., 2001; 61: 6213-6218.
36 Ray DM, Bernstein SH, Phipps RP. Human multiple myeloma cells express peroxisome proliferator-activated receptor gamma and undergo apoptosis upon exposure to PPARgamma ligands. Clin Immunol. 2004 Nov; 113(2):203-213.
37 董育玮，王兴鹏，吴恺et al．过氧化物酶增殖物活化受体γ在胰腺癌生长中的调节作用．中华内科杂志，2003，42(7)：479-482．
38 Zhu L, Gong B, Bisgaier CL, et al. Induction of PPARgamma expression in human THP-1 monoeytic leukemia cells by 9-cis-retinoic acid is associated with cellular growth suppression.Biochm Biophys Res Commun. 1998 Oct 29; 251(3): 842-848.
39 LIUH, ZANGC,FENNERMH.PPARγand ATKA inhibit the invasion of huaman breast cancer cells in vitro [J]. Breast cancer Rsearch and Treatment, 2003, 79: 63-67.
40 周亭芳，庄英帜，曹建国．罗格列酮增强顺铂抑制入肺腺癌细胞增殖作用[J]．中国药理学通报，2005，21(1)：88-91．
2. ZhaoX, Demary K, WongL, et al. Retinoic acid receptor-independent mechanism of apoptosis of melanoma cells by the retinoid CD437(AHPN) [J]. Cell Death Differ,2001;8(9):878-886.
3. Kliewer SA, Umesono K,Noonan DJ, et al. Convergence of 9-cis retinoic acid and peroxisome proliferator signaling pathways through heterodimer formation of their receptors Nature. 1992 Aug 27;358(6389):771-774.
4. Marcus SL,Capone JP,Rachubinski RA.Identification of COUP-TFII as a Peroxisome proliferator response element binding factor using genetic seletion I yeast: COUP-TFII activates transcription in yeast but antagonizes PPAR signaling in mammalian cells. Mol Cell Endocrinol 1996, 120: 31- 39.
5. Rao MS, Reddy JK. Peroxisomal beta-oxidation and steatohepatitis. Semin Liver Dis. 2001; 21(1):43-55.
6. Daynes RA,Jones DC.Emerging roles of PPARs in inflammation and immunity.Nat Rev Immunol.2002 Oct;2(10):748-759.
7. Miyahara T,Schrum L,Rippe R,et al. Peroxisome proliferator-activated receptors and hepatic stellate cell activation.J Biol Chem.2000 Nov 17;275(46):35715-35722.
8. Scheen AJ, Paquot N. PPAR-gamma receptors, new therapeutic target in metabolic and cardiovascular diseasesRev Med Liege. 2005 Feb; 60(2):89-95.
9. Martens FM, Visseren FL, Lemay J, et al. Metabolic and additional vascular effects of thiazolidinediones. Drugs. 2002; 62(10): 1463-1480.
10. Shouwei Han, Neil Sidell, Paul B, et al. Up-regulation of p21 Gene Expression by Peroxisome Proliferator-Activated Receptorγ in Human Lung Carcinoma Cells. Clin Cancer Res. 2004, Mar 15; 10(6):1911-1919.
11. Reed JC. Apoptosis mechanisms: implications for cancer drug discovery.??Oncology (Huntingt). 2004 Nov; 18(13 Suppl 10):11-20.
12. Yoshimura R, Matsuyama M, Segawa Y et al. Expression of peroxisome proliferator-activated receptors (PPARs) in human urinary bladder carcinoma and growth inhibition by its agonists. Int J Cancer. 2003 May 1; 104(5): 5 97-602.
13. Guan YF, Zhang YH, Breyer RM et al. Expression of peroxisome proliferator-activated receptor gamma (PPARgamma) in human transitional bladder cancer and its role in inducing cell death. Neoplasia. 1999 Oct;1(4):330-339.
14. Heaney AP, Fernando M, Melmed S. PPAR-gamma receptor ligands: novel therapy for pituitary adenomas. J Clin Invest. 2003 May; 111(9): 1381-8.
15. Keshamouni VG, Arenberg DA, Reddy RC, et al. PPAR-gamma activation inhibits angiogenesis by blocking ELR+CXC chemokine production in non-small cell lung cancer. Neoplasia. 2005 Mar;7(3):294-301
16. Lee KS, Park SJ, Hwang PH, et al. PPAR-gamma modulates allergic inflammation through up-regulation of PTEN. FASEBJ.2005 Jun; 19(8): 1033-5.
17. Konturek PC, Kania J, Konturek JW, et al. H. pylori infection, atrophic gastritis, cytokines, gastrin, COX-2, PPAR gamma and impaired apoptosis in gastric carcinogenesis. Med Sci Monk. 2003 Jul; 9(7):SR53-66.
18. Guan Y. Targeting peroxisome proliferator-activated receptors (PPARs) in kidney and urologic disease. Minerva Urol Nefrol. 2002 Jun; 54(2):65-79.
19. Wijnhoven BP, Lindstedt EW, Abbou M et alJ Dinjenens, WN.Molecular genetic analysis of the von Hippel—Lindau and human Peroxisome Proliferator-activated receptor gamma tumor-suppressor genes in adenocarcinomas of the gastroesophageal junction. Int J Cancer, 2001; 94(6): 891—895
20. Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of peroxisome proliferator-activated receptor gamma in non-small cell lung Cancer.Cancer Res, 2000; 60(4): 1129-1138.
21. HeTC,ChanTA,VogelsteinB, et al. PPARγis an APC-Regulated Target of??Nonsteroidal Anti-InflammatoryDrugs. Cell, 1999: 336～345.
22. Smith SA. Peroxisome proliferator-activated receptors and the regulation of mammalian lipid metabolism. Biochem Soc Trans. 2002 Nov; 30(Pt 6): 1086-1090.
23. Altiok S, Xu M, Spiegelman BM. PPARgamma induces cell cycle withdrawal: inhibition of E2F/DP DNA-binding activity via down-regulation of PP2A. Genes Dev. 1997Aug 1; 11(15): 1987-1998.
24. Tsubouchi Y, Sano H, Kawahito Y et al. Inhibition of human lung cancer cell growth by the peroxisome proliferator-activated receptor-gamma agonists through induction of apoptosis. Biochem Biophys Res Commun. 2000 Apr 13; 270(2):400-405.
25．周亭芳，庄英帜，曹建国．罗格列酮增强顺铂抑制人肺腺癌细胞增殖作用[J]中国药理学通报，2005，21(1)：88-91．
26. Sherr CJ. The Pezzoller Lecture: cancer cell cycle revisited. Cancer Res, 200; 60: 3689.
27. Koga H, Sakisaka S, Harada M, et al. Involvement of p21(WAF1/Cip1), p27(Kip1), and p18(INK4c) in troglitazoneinduced cell-cycle arrest in human hepatoma cell lines. Hepatology 2001; 33: 1087-1097.
28. Itami A, Watanabe G, Shimada Y, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth of pancreatic cancers both in vitro and in vivo. Int J Cancer 2001; 94: 370-376.
29. Motomura W, Okumura T, Takahashi N, et al. 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: 5558-5564.
30. Yin F, Wakino S, Liu Z, et al. Troglitazone inhibits growth of MCF-7 breast carcinoma cells by targeting G1 cell-cycle regulators. Bioehem Biophys Res Commun 2001; 286: 916-922.
31. Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of peroxisome proliferator-activated receptor gamma in nonsmall-cell lung cancer.??Cancer Res 2000; 60:1129-1138.
32. Morrison RF, Farmer SR. Role of PPARgamma in regulating a cascade expression of cyclin-dependent kinase inhibitors, p18(INK4c) and p21(Waf1/Cip1), during adipogenesis. J Biol Chem 1999; 274:17088-17097.
33. Elnemr A, Ohta T, Iwata K, et al. PPARgamma ligand (thiazolidinedione) induces growth arrest and differentiation markers of human pancreatic cancer cells. Int J Oncol 2000; 17:1157-1164.
34. Itami A, Watanabe G, Shimada Y, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth of pancreatic cancers both in vitro and in vivo. Int J Cancer 2001; 94: 370-376.
35. Tien HF, Wang CH. CD7 positive hematopoietic progenitors and acute myeloid leukemia and other minimally differentiated leukemia. Leuk Lymphoma. 1998 Sep; 31(1-2):93-98.
36. samura RY, Egashira N, Miyai Set al.Molecular pathology of the pituitary. Development and functional differentiation of pituitary adenomas. Front Horm Res. 2004; 2:20-33.
37. Asou H, Verbeek W, Williamson E, et al. Growth inhibition of myeloid leukemia cells by troglitazone, a ligand for peroxisome proliferator activated receptor gamma, and retinoids. Int J Oncol 1999; 15:1027-1031.
38. Ray DM, Bernstein SH, Phipps RP. Human multiple myeloma cells express peroxisome proliferator-activated receptor gamma and undergo apoptosis upon exposure to PPARgamma ligands. Clin Immunol. 2004 Nov; 113(2):203-213.
39．董育玮，王兴鹏，吴恺 et al．过氧化物酶增殖物活化受体γ在胰腺癌生长中的调节作用．中华内科杂志，2003，42(7)：479-482．
40. Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of peroxisome proliferator-activated receptor gamma in non-small cell lung Cancer. Cancer Res, 2000; 60(4): 1129-1138.
41．曾益新编(主编)肿瘤学，第三版．北京：人民卫生出版社，2003：201．
42. Pitot HC, Glauert HP, Hanigan M. The significance of selected biochemical markers in the characterization of putative initiated cell populations in rodent liver.??Cancer Lett. 1985 Oct; 29(1):1-14.
43．秦锁富，查锡良，陈惠黎．亚硒酸钠对人肝癌细胞株甲胎蛋白纤维连接蛋白及细胞膜糖链的影响[J]．肿瘤，1991，11(3)：133．
44. Cox PJ, Ryan DA, Hollis FJ, et al. Absorption, disposition, and metabolism of rosiglitazone, a potent thiazolidinedione insulin sensitizer, in humans. Drug Metab Dispos, 2000; 28(7): 772-780.1. Michalik L, Wahli W. Peroxisome proliferator-activated receptors:three isotypes for a multitude of functions. [J]. Curr Opin Biotechnol. 1999 Dec; 10(6):564-570.
2. Issemann I, Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. [J]. Nature. 1990 Oct 18; 347(6294):645-650.
3 Farrow B, Evers BM. Activation of PPARgamma increases PTEN expression in??pancreatic cancer cells. (J) . Biochem Biophys Res Commun. 2003 Jan 31; 301(1):50-53.
4 Berger J, Moller DE. The mechanisms of action of PPARs. [J]. Annu Rev Med. 2002; 53: 409-435.
5 Subbaramaiah K, Lin DT, Hart JC,et al. Peroxisomeproliferator-actived receptor gamma ligands suppress the transcriptional activation of cyclooxygenase-2. Evidence for involvement of activator protein-1 and CREB-binding protein/p300.J Biol Chem. 2001 Apr 13; 276(15): 12440-12778. Epub 2001 Jan 23.
6 Zander T, Kraus JA, Grommes C, et al. Induction of apoptosis in human and rat glioma by agonists of the nuclear receptor PPARgamma. J Neurochem 2002; 81: 1052-1060.
7 Chattopadhyay N, Singh DP, Heese O, et al. Expression of peroxisome proliferator-activated receptors (PPARS) in human astrocytic cells: PPARgamma agonists as inducers of apoptosis.J Neurosci Res 2000; 61: 61-74.
8 Yoshida K, Tanabe K, Fujii D, et al.Induction mechanism of apoptosis by troglitazone through peroxisome proliferator-activated receptor-gamma in gastric carcinoma cells. [J]. Anticancer Res. 2003 Jan-Feb; 23(1A):267-273.
9 Shimada T, Kojima K, Yoshiura K, et al. Characteristics of the peroxisome proliferator-activated receptor gamma (PPARgamma)ligand-induced apoptosis in colon cancer cells. Gut 2002; 50: 658-664.
10 Eibl G, Wente MN, Reber HA, Hines OJ. Peroxisome proliferatoractivated receptor gamma induces pancreatic cancer cell apoptosis. Biochem Biophys Res Commun 2001; 287: 522-529.
11 Toyoda M, Takagi H, Horiguchi N, et al. A ligand for peroxisome proliferator-activated receptor gamma inhibits cell growth and induces apoptosis in human liver cancer cells. Gut 2002; 50: 563-567.
12 Ohta K, Endo T, Haraguchi K, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth and induce apoptosis of human papillary thyroid carcinoma cells. J Clin Endocrinol Metab 2001; 86: 2170-2177.
13 Tontonoz P, Hu E, Spiegelman BM. Stimulation of adipogenesis in fibroblasts by??PPAR gamma 2, a lipid-activated transcription factor. Cell 1994; 79: 1147-1156.
14 Okuno A, Tamemoto H, Tobe K, et al. Troglitazone increases the number of small adipocytes without the change of white adipose tissue mass in obese Zucker rats. J Clin Invest 1998; 101: 1354-1361.
15 Tontonoz P, Singer S, Forman BM, et al. Terminal differentiation of human liposarcoma cells induced by ligands for peroxisome proliferator-activated receptor gamma and the retinoid X receptor.Proc Natl Acad Sci USA 1997; 94: 237-241.
16 Elnemr A, Ohta T, Iwata K, et al. PPARgamma ligand (thiazolidinedione) induces growth arrest and differentiation markers of human pancreatic cancer cells. Int J Oncol 2000; 17: 1157-1164.
17 Altiok S, Xu M, Spiegelman BM. PPARgamma induces cell cycle withdrawal: inhibition of E2F/DP DNA-binding activity via down-regulation of PP2A. Genes Dev. 1997 Aug 1; 11(15): 1987-1998.
18 Morrison RF, Farmer SR. Role of PPARgamma in regulating a cascade expression of cyclin-dependent kinase inhibitors, p18(INK4c) and p21(Waf1/Cip1), during adipogenesis. J Biol Chem 1999; 274: 17088-97.
19 Koga H, Sakisaka S, Harada M, et al. Involvement of p21(WAF1/Cip1), p27(Kip1), and p18(INK4c) in troglitazoneinduced cell-cycle arrest in human hepatoma cell lines. Hepatology2001; 33: 1087-1097.
20 Itami A, Watanabe G, Shimada Y, et al. Ligands for peroxisome proliferator-activated receptor gamma inhibit growth of pancreatic cancers both in vitro and in vivo. Int J Cancer 2001; 94: 370-376.
21 Motomura W, Okumura T, Takahashi N, et al. 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: 5558-5564.
22 Yin F, Wakino S, Liu Z, et al. Troglitazone inhibits growth of MCF-7 breast carcinoma cells by targeting G1 cell-cycle regulators. Biochem Biophys Res Commun 2001; 286: 916-922.
23 Chang TH, Szabo E. Induction of differentiation and apoptosis by ligands of??peroxisome proliferator-activated receptor gamma in nonsmall-cell lung cancer. Cancer Res 2000; 60: 1129-1138.
24 XinX, Yang S, Kowalski J, Gerritsen ME. Peroxisome proliferatoractivated receptor gamma ligands are potent inhibitors of angiogenesis in vitro and in vivo. J Biol Chem 1999; 274: 9116-9121.
25 Panigrahy D, Singer S, Shen LQ, et al. PPARgamma ligands inhibit primary-tumor growth and metastasis by inhibiting angiogenesis. J Clin Invest 2002; 110: 923-932.
26 Panigrahy D, Dulak J, Piatkowska E, et al. Ligands of peroxisome proliferator-activated receptor-gamma increase the generation of vascular endothelial growth factor in vascular smooth-muscle cells and in macrophages. Acta Biochim Pol 2000; 47: 1147-1157.
27 Benjamin L, Golijanin D, Itin A et al. Selective ablation of immature blood vessels in established tumours follows vascular endothelial growth factor withdrawal. J Clin Invest 1999; 103: 159-165.
28 Freed MI, Allen A, Jorkasky DK, DiCicco RA. Systemic exposure to rosiglitazone is unaltered by food. Eur J Clin Pharmacol 1999;55: 53-56.
29 Sierra-Honigmann MR, Nath AK, Murakami C et al. Biological action of leptin as an angiogenic factor. Science 1998; 281: 1683- 1686.
30 Goetze S, Bungenstock A, Czupalla C, et al. Leptin induces endothelial-cell migration through Akt, which is inhibited by PPARgamma-ligands. Hypertension 2002; 40: 748-754.
31 Cui Y., Miyoshi K., Claudio E., Siebenlist U. K., Gonzalez F. J., Flaws J., Wagner K-U., Hennighausen L. Loss of the peroxisome proliferation-activated receptor gamma (PPAR(?)) does not affect mammary development and propensity for tumor formation but leads to reduce fertility. J. Biol. Chem., 277: 17830-17835, 2002.
32 Takayuki Ikezoe, Carl W, et al. Mutational Analysis of the Peroxisome Proliferator-activated Receptor (?) Gene in Human Malignancies. Cancer Research 61, 2001, July 1, 5307-5310.
33 Palakurthi S. S., Aktas H., Grubissich L. M., Mortensen R. M., Halperin J. A. ??Anticancer effects of thiazolidinediones are independent of peroxisome proliferator-activated receptor γand mediated by inhibition of translation initiation. Cancer Res., 61: 6213-6218, 2001.
34 Rossi A., Kapahi P., Natoli G, Takahashi T., Chen Y., Karin M., Santoro M. G. Anti-inflammatory cyclopentenone prostaglandins are direct inhibitors of INB kinase. Nature (Lond.), 403: 103-106, 2000.
35 Palakurthi S. S., Aktas H., Grubissich L. M., Mortensen R. M., Halperin J. A. Anticancer effects of thiazolidinediones are independent of peroxisome proliferator-activated receptor γand mediated by inhibition of translation initiation. Cancer Res., 2001; 61: 6213-6218.
36 Ray DM, Bernstein SH, Phipps RP. Human multiple myeloma cells express peroxisome proliferator-activated receptor gamma and undergo apoptosis upon exposure to PPARgamma ligands. Clin Immunol. 2004 Nov; 113(2):203-213.
37 董育玮，王兴鹏，吴恺et al．过氧化物酶增殖物活化受体γ在胰腺癌生长中的调节作用．中华内科杂志，2003，42(7)：479-482．
38 Zhu L, Gong B, Bisgaier CL, et al. Induction of PPARgamma expression in human THP-1 monoeytic leukemia cells by 9-cis-retinoic acid is associated with cellular growth suppression.Biochm Biophys Res Commun. 1998 Oct 29; 251(3): 842-848.
39 LIUH, ZANGC,FENNERMH.PPARγand ATKA inhibit the invasion of huaman breast cancer cells in vitro [J]. Breast cancer Rsearch and Treatment, 2003, 79: 63-67.
40 周亭芳，庄英帜，曹建国．罗格列酮增强顺铂抑制入肺腺癌细胞增殖作用[J]．中国药理学通报，2005，21(1)：88-91．