The effects of peroxisome proliferator-activated receptor gamma agonists and fluorouracil on Colon 38 cancer growth in vitro

• Authors: Karolina Beda-Maluga , Julita Fuss-Chmielewska , Jacek Świętosławski , Katarzyna Winczyk

Title variants

PL

Wpływ agonistów receptorów aktywowanych proliferatorami peroksysomów typu gamma i fluorouracylu na wzrost komórek raka linii Colon 38 in vitro

• Languages of publication: EN

Abstracts

EN

Introduction: Colon cancer is a very serious medical problem in Europe and also in Poland. For many years fluorouracil has been used as a main agent in chemotherapy, but its effectiveness is not sufficient. Recently several modern drugs were introduced in the treatment of colon cancer and also various of therapeutic options have been tested but the survival time of patients with cancer has not been extended significantly. For that reason more efficient drugs have still been looked for. One of the potential anticancer drugs are thiazolidinediones (TZDs) – agonists of peroxisome proliferator-activated receptors gamma (PPARγ) which in the last years were used in diabetes treatment. Therefore, we decided to examine the effect of two TZDs – pioglitazone (PIO) and rosiglitazone (ROS) on the growth of murine colon cancer and to compare their action with the efficacy of routinely used fluorouracil (FU). Furthermore, we evaluated the PPARγ expression in colon cancer cells by using the immunocytochemical method. Material and methods: Cell line of murine cancer – Colon 38 was used in our experiment. The growth of cancer cells was assessed by using EZ4Y kit based on the modified colorimetric Mosmann method. In 24 and 48 h cell culture the effects of ROS and PIO at concentrations 10-4, 10-5, 10-6, 10-7 M and FU at concentrations 4 x 10-6 and 10-6 M were examined. Immunochistochemistry was performed with the use of murine specific polyclonal antibodies anti-PPARγ1,2 and the streptavidin-biotin-peroxidase method. Results: The immunopositive reaction for PPARγ was shown in the nuclei of Colon 38 cells. Both, the examined TZDs and fluorouracil significantly decreased the growth of colon cancer and their efficacy was dependent on the concentration of examined compounds and also the incubation time. Rosiglitazone, in all used concentrations, acted more strongly than PIO. Fluorouracil showed anti-cancer activity only in 48 h culture and its inhibitory effect was weaker than both TZDs at the highest concentration (10-4 M). Conclusions: Our data indicate that PPARγ agonists, especially rosiglitazone, inhibit the growth of Colon 38 cancer. However, the potential usefulness of rosiglitazone and pioglitazone for the chemoprevention and treatment of colon cancer requires further studies.

PL

Wstęp: Rak jelita grubego stanowi poważny problem medyczny zarówno w Polsce, jak i w całej Europie. Przez wiele lat jako główny chemioterapeutyk stosowano fluorouracyl, niestety jego skuteczność okazała się niewystarczająca. Zastosowanie w terapii raka jelita grubego nowych leków i różnych opcji terapeutycznych nie wydłużyło istotnie czasu przeżycia pacjentów. Z tego powodu nadal poszukiwane są bardziej skuteczne formy leczenia. Związkami o potencjalnym działaniu przeciwnowotworowym są tiazolidinediony (TZDs) – agoniści receptorów gamma aktywowanych proliferatorami peroksysomów (PPARγ), które w ostatnich latach wykorzystywano w leczeniu cukrzycy. W związku z tym, w pracy oceniono wpływ dwóch TZDs – pioglitazonu (PIO) i roziglitazonu (ROS) na wzrost mysiego raka jelita grubego, a ich działanie porównano ze skutecznością fluorouracylu (FU). Ponadto w komórkach raka jelita grubego metodą immunohistochemiczną oceniono ekspresję PPARγ. Materiał i metody: Badanie przeprowadzono na linii komórkowej mysiego raka jelita grubego – Colon 38. Wzrost komórek nowotworowych mierzono za pomocą zestawu EZ4Y opartego na metodzie kolorymetrycznej Mosmann’a. W hodowli 24- i 48-godzinnej oceniono wpływ ROS i PIO w stężeniach 10-4, 10-5, 10-6, 10-7 M, a działanie fluorouracylu w stężeniach 4 x 10-6 i 10-6 M. W badaniu immunohistochemicznym wykorzystano mysie poliklonalne przeciwciała anty-PPARγ1,2 i streptawidynowo-biotynowo-peroksydazową metodę wizualizacji. Wyniki: W komórkach raka Colon 38 wykazano immunopozytywny jądrowy odczyn dla PPARγ. Badane TZDs i fluorouracyl znacząco zahamowały wzrost raka jelita grubego, a ich skuteczność zależna była od stężenia i czasu inkubacji. Działanie ROS było silniejsze niż PIO we wszystkich użytych stężeniach. Przeciwnowotworowe działanie fluorouracylu stwierdzono po 48 godzinach inkubacji, jednakże jego efekt był słabszy niż wpływ TZDs w najwyższym badanym stężeniu (10-4 M). Wnioski: Uzyskane wyniki wskazują, że agoniści receptorów PPARγ, a w szczególności roziglitazon, hamują wzrost raka Colon 38. Jednakże wykorzystanie roziglitazonu i pioglitazonu w leczeniu raka jelita grubego wymaga dalszych badań.

Keywords

EN

PPARγ agonists; agoniści PPARγ; colon cancer; fluorouracil; fluorouracyl; pioglitazon; pioglitazone; rak jelita grubego; rosiglitazone; roziglitazon

Discipline

• MEDICINE: MEDICINE

• Publisher: Łódzkie Towarzystwo Naukowe
• Journal: Folia Medica Lodziensia
• Year: 2011
• Volume: 38
• Issue: 2
• Pages: 227-243

Contributors

author

Karolina Beda-Maluga

• Department of Neuroendocrinology, Chair of Laboratory Diagnostics, Medical University of Lodz, Poland

author

Julita Fuss-Chmielewska

• Department of Neuroendocrinology, Chair of Laboratory Diagnostics, Medical University of Lodz, Poland

author

Jacek Świętosławski

• Department of Neuroendocrinology, Chair of Laboratory Diagnostics, Medical University of Lodz, Poland

author

Katarzyna Winczyk

• Department of Neuroendocrinology, Chair of Laboratory Diagnostics, Medical University of Lodz, Poland

References

• Kulbacka J, Saczko J, Chwiłkowska A. Rak jelita grubego – charakterystyka i oporność na leczenie. Onkol Prak Klin. 2008; 4: 135–140.
• Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics, 2009. CA Cancer J Clin. 2009; 59: 225-249.
• Schrag D. The price tag on progress – chemotherapy for colorectal cancer. N Engl J Med. 2004; 351: 317-319.
• Hurwitz H, Fehrenbacher L, Novotny W, Cartwright T, Hainsworth J, Heim W et al. Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N Engl J Med. 2004; 350: 2335-2342.
• Iqbal S, Lenz H-J. Integration of novel agents in the treatment of colorectal cancer. Cancer Chemother Pharmacol. 2004; 54: S32-39.
• Saltz L. Current role of bevacizumab in colorectal cancer. Clin Adv Hematol Oncol. 2009; 7: 375-376
• Marshall JL, Gramont A, Köhne CH. Accomplishments in 2008 in the adjuvant treatment of colon cancer. Gastrointest Cancer Res. 2009; 3: 2–7.
• Schoonjans K, Staels B, Auwerx J. The peroxisome proliferator activated receptors (PPARs) and their effects on lipid metabolism and adipocyte differentiation. Biochim Biophys Acta. 1996; 1302: 93-109.
• Desvergne B, Wahli W. Peroxisome proliferator-activated receptors: nuclear control of metabolizm. Endocr Rev. 1999; 20: 649-688.
• Kersten S, Desvergne B, Wahli W. Roles of PPARs in health and disease. Nature. 2000; 405: 421-425.
• Koeffer HP. Peroxisome proliferator-activated receptor γ and cancers. Clin Cancer Res. 2003; 9: 1-9.
• Winczyk K. Receptory aktywowane proliferatorami peroksysomów gamma - ich znaczenie w karcinogenezie i terapii nowotworów. Fol Med Lodz. 2007; 34: 117-140.
• Winczyk K. The role of peroxisome proliferators-activated receptors (PPARgamma) in neoplasms of endocrine glands. Pol J Endocrinol. 2008; 59:156-166.
• Blanquicett C, Roman J, Hart M. Thiazolidinediones as anti-cancer agents. Cancer Ther. 2008; 6: 25–34.
• Wang YL, Miao Q. To live or to die: prosurvival activity of PPAR γ on cancers. PPAR Res. doi:10.1155/2008/209629.
• Hatton JL, Yee LD. Clinical Use of PPARgamma Ligands in Cancer. PPAR Res. 2008:159415. Dec 18.
• Thompson EA. PPAR γ physiology and pathology in gastrointestinal ephithelial cells. Mol Cells. 2007; 24: 167-176.
• ShimadaT, Kojima K, Yoshiura K, Hiraishi H, Terano A. Characteristics of the peroxisome proliferators-activated receptor gamma ligand induced apoptosis in colon cancer cells. Gut. 2002; 50: 658-664.
• Osawa E, Nakajima A, Wada K, Ishimine S, Fujisawa N, Kawamori T et al. Peroxisome proliferators-activated receptor gamma ligands suppress colon carcinogenesis induced by azoxymethane in mice. Gastroenterology. 2003; 124: 361-367.
• Yasui Y, Hosokawa M, Kohno H, Tanaka T, Miyashita K. Troglitazone and 9cis, 11trans, 13trans-conjugated linolenic acid: comparison of their antiproliferative and apoptosis-inducing effects on different colon cancer cell lines. Chemotherapy. 2006; 52: 220-225.
• Chintharlapalli S, Smith R, Samudio I, Zhang W, Safe S. 1, 1-bis(3’-indolyl)-1-(psubstitutedphenyl) methanes induce peroxisome proliferators – activated receptor γ–mediated growth inhibition, transactivation, and differentiation markers in colon cancer cells. Canc Res. 2004; 64: 5994-6001.
• Lee CJ, Han JS, Seo CY, Park TH, Kwon HC, Jeong JS et al. Pioglitazone, a synthetic ligand for PPARgamma, induces apoptosis in RB-deficient human colorectal cancer cells. Apoptosis. 2006; 11: 401-411.
• Zhang YQ, Tang XQ, Sun L, Dong L, Qin Y, Liu HQ et al. Rosiglitazone enhances fluorouracil - induced apoptosis of HT-29 cells by activating peroxisome proliferator - activated receptor gamma. World J Gastroenterol. 2007; 13: 1534-1540.
• Shen D, Deng C, Zhang M. Peroxisome proliferator-activated receptor gamma agonists inhibit the proliferation and invasion of human colon cancer cells. Postgrad Med J. 2007; 83: 414-419.
• Corbett TH, Griswold DP, Roberts BJ, Peckham JC, Schabel FM. Tumor induction relationships in development of transplantable cancers of the colon in mice for chemotherapy assays, with a note on carcinogen structure. Cancer Res. 1975; 35: 2434-2439.
• Pajtasz-Piasecka E, Glazman-Kuśnierczyk, H, Salwa J, Konarski L, Radzikowski C. Growth inhibition of transplantable tumors in mice by mIL 2 secreting murine plasmocytoma cells used alone or in combination with a cytostatic agent. Arch Immunol Ther Exp. 1995; 43: 281-292.
• 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: 1046–1052.
• Lin MS, Chen WC, Bai X, Wang YD. Activation of peroxisome proliferator-activated receptor gamma inhibits cell growth via apoptosis and arrest of the cell cycle in human colorectal cancer. J Dig Dis. 2007; 8: 82-88.
• Takano S, Kubota T, Nishibori H, Hasegawa H, Ishii Y, Nitori N et al. Pioglitazone, a ligand for peroxisome prolife as an inhibitor of colon cancer liver metastasis. Anticancer Res. 2008; 28: 3593-3599.rator-activated receptor-gamma acts
• Fu JN, Li J, Tan Q, Yin HW, Xiong K, Wang TY et al. Thioredxin reductase inhibitor ethaselen increases the drug sensitivity of the colon cancer cell line LoVo towards cisplatin via regulation of G1 phase and reversal of G2/M phase arrest. Invest New Drugs. 2011; 29: 627-636.
• Mełeń-Mucha G, Winczyk K, Pawlikowski M. Effects of somatostatin analogs octreotide and lanreotide on the proliferation and apoptosis in colon 38 tumor: interaction with 5-fluorouracil. Neuroendocrinol Lett. 2000; 21: 137-142.
• Winczyk K, Pawlikowski M, Karasek M. Melatonin and RZR/ROR receptor ligand CGP 52608 induce apoptosis in the murine colon cancer. J Pineal Res. 2001; 31: 179-182.
• Mełeń-Mucha G, Ławnicka H, Mucha S. Ten combine effects of progluminde and fluorouracil on the growth of murine Colon 38 cancer cells in vitro. Pol J Endocrinol. 2005; 6: 933-938.
• Winczyk K, Fuss-Chmielewska J, Lawnicka H, Pawlikowski M, Karasek M. Luzindole but not 4-phenyl-2- propionamidotetralin (4P-PDOT) diminishes the inhibitory effect of melatonin on murine Colon 38 cancer growth in vitro. Neuro Endocrinol Lett. 2009; 30: 657-662.
• Ferruzzi P, Ceni E, Tarocchi M, Grappone C, Milani S, Galli A et al. Thiazolidinediones inhibit growth and invasiveness of the human adrenocortical cancer cell line H295R. J Clin Endocrinol. 2005; 90: 1332-1339.
• Choi IK, Kim YH, Kim JS, Seo JH. PPAR-gamma ligand promotes the growth of APC-mutated HT-29 human colon cancer cells in vitro and in vivo. Invest New Drugs. 2008; 26: 283-288.
• Mełeń-Mucha G, Ławnicka H. Lepton promotes the growth of Colon 38 cancer cells and interferes with the cytotoxic effect of fluorouracil in vitro. Pol J Endocrinol. 2007; 1: 2-6.
• Lago RM, Singh PP, Nesto RW. Congestive heart failure and cardiovascular death in patients with prediabetes and type 2 diabetes given thiazolidinediones: a meta-analysis of randomised clinical trials. Lancet. 2007; 29: 1129-1136.
• Mannucci E, Monami M, Di Bari M, Lamanna C, Gori F, Gensini GF et al. Cardiac safety profile of rosiglitazone: a comprehensive meta-analysis of randomized clinical trials. Int J Cardiol. 2010; 143: 135-140.
• Komajda M, McMurray JJ, Beck-Nielsen H, Gomis R, Hanefeld M, Pocock SJ et al. Heart failure events with rosiglitazone in type 2 diabetes: data from the RECORD clinical trial. Eur Heart J. 2010; 31: 824-831.
• Woodcock J, Sharfstein JM, Hamburg M. Regulatory action on rosiglitazone by the U.S. Food and Drug Administration. N Engl J Med. 2010; 36: 1489-1491.
• Blind E, Dunder K, de Graeff PA, Abadie E. Rosiglitazone: a European regulatory perspective. Diabetologia. 2011; 54: 213-218.

• Document Type: paper