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MCF-7 İnsan Meme Kanseri Hücre Soyunda Doksorubisin Öncesi ve Sonrası GST İzozimlerinin, İlaç Dirençlilik Proteinlerinin ve Apoptotik Etkisinin Araştırılması

Yıl 2020, Sayı: 10, 1 - 18, 17.04.2020
https://doi.org/10.38079/igusabder.631695

Öz

Amaç: Kanser hastalığının tedavisinde karşılaşılan klinik sorunlardan biri hastalara uygulanan kemoterapiye karşı tümör hücrelerinin geliştirdiği dirençtir. Tümör hücrelerinin ilaçlara karşı gösterdiği direncin asıl kaynaklarından biri, ilaçların hücre dışına atılmasını sağlayan membran proteinlerinin en önemli üyelerinden ABC (ATP-binding cassette) taşıyıcı proteinleridir. İlaç dirençlilik proteinlerinin yanında diğer hücre içi proteinlerinde etkin olabileceği bilinmektedir. Bu bağlamda, alkilleyici özellikteki kanser ilaçlarına gelişen dirençte, hücre içi glutatyon ve glutatyon S- konjugatlarının seviyelerinin artmasının rolünün olduğu bildirilmiştir. Bu çalışmada, Michigan Cancer Foundation-7 (MCF-7) meme kanseri hücre hattında Glutatyon S-transferaz (GST) izozimlerinin çoklu ilaç direnç mekanizmasındaki bazı önemli proteinlerin doksorubisin uygulamasıyla ilişkilerinin incelenmesi amaçlanmıştır.
Yöntem: Bu çalışmada ilaç uygulanmış ve uygulanmamış meme kanserli MCF-7 hücre hattında GST enzim ailesi ve ABC taşıyıcı proteinlerin ekspresyon ifadeleri immünositokimya yöntemiyle incelenmiştir.
Bulgular: Bu çalışmada, ilaç uygulanmış hücre hatlarında GSTP1, GSTT1, GSTM1, GSTA1, GSTO1, GSTZ1 ve GSTK1 protein ifadelerinin ilaç uygulanmamış hücrelere oranla daha yüksek olduğu görülmüştür; GSTS1 proteini kontrol ve deney gruplarının ikisinde de tespit edilememiştir. İlaç uygulanmış MCF-7 hücre hattında MRP (Multidrug resistance-associated) 2,3,6,7 protein ifadelerinin ilaç uygulanmamış hücrelere oranla daha fazla olduğu görülmüştür. MDR1 (multidrug resistance protein) ve MRP1 proteinleri izlenememiştir. İlaç uygulanmış meme kanserli MCF-7 hücre hattında Bcl-2, p53, p38, caspase-3 protein ifadelerinin ilaç uygulanmamış hücrelere oranla daha fazla olduğu görülmüştür.
Sonuç: ABC süper ailesi üyelerinden MRP 2,3,6 ve 7 ile Faz II enzimleri arasında bulunan GSTP1, GSTT1, GSTM1, GSTA1, GSTO1, GSTZ1 ve GSTK1 izozimlerinin, MCF-7 kanser hücre hattında doksorubisine karşı oluşan ilaç dirençliliğinde rolleri olduğu belirlenmiştir. 

Destekleyen Kurum

KIRIKKALE ÜNİVERSİTESİ BAP BİRİMİ

Proje Numarası

2015/125

Teşekkür

KIRIKKALE ÜNİVERSİTESİ BAP BİRİMİNE TEŞEKKÜR EDERİZ.

Kaynakça

  • Tozkoparan B, Peri Aytaç S. Kanser kemoterapisinde terapötik hedef olarak glutatyon S-transferazlar. Hacettepe Üniversitesi, Eczacılık Fakültesi Dergisi. 2007;7(2):139-164.
  • Dünyada Kanser İstatistikleri. Türk Kanser Araştırma ve Savaş Kurumu (TKASKD).http://turkkanser.org.tr. Yayınlanma tarihi 1995. Erişim tarihi 20.09.2019.
  • Onur H. Tıbbi Onkoloji Kitabı. Ankara Üniversitesi Tıp Fakültesi: Antıp Yayınları; 2005.
  • Gate L, Tew KD. Glutathione S-transferases as emerging targets, Expert Opin Ther. Targets. 2001;5(4):477-489.
  • Szakács G, Paterson, JK, Ludwig JA, Booth-Genthe C, Gottesman MM. Targeting multidrug resistance in cancer. Nat Rev Drug Discov. 2006;5(3):219-34.
  • Stavrovskaya AA, Stromskaya TP. Transport proteins of the ABC familya and multidrug resistance of tumor cells. Biochemistry (Mosc). 2008;73(5):592-604.
  • Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics. CA Cancer J Clin 2009;59(4):225-249. doi: 10.3322/caac.20006.
  • Tuncer M. Significance of cancer in Turkey, the burden of disease and cancer control policies. Cancer Control in Turkey, Ankara, Onur Press, Health Ministry Publication. 2008:74;5-9.
  • Marie JP, Legrand O. Drug resistance in acute leukaemia and reversion. Turk J Med Sci. 2003;33:271-279.
  • Turgut S, Yaren A, Kursunluoğlu R, Turgut G. MDR1 C3435T polymorphism in patients with breast cancer. Arch Med Res. 2007:38;539-544.
  • Eaton DL, Bammler TK. Concise Review of glutathione s-transferase and their significance to toxicology. Toxicological Sciences. 1999;49:156-164.
  • Oguztuzun S, Abu-Hijleh A, Coban T, et al. GST isoenzymes in matched normal and neoplastic breast tissue. Neoplasma. 2011;58(4):304-310.
  • Buller Al, Clapper M, Smith T, Tew KD Glutathione S-trasnferase in nitrogen mustard-resistance and-sensitive. Molecular Pharmacology. 1987;31(6):575-578.
  • Oguri T, Fujimara Y, Katoh O, et al. Glutathione S-transferase gene expression and platinum drug exposure in human lung cancer. Cancer Letters. 2000;156:93-99.
  • Ban N, Takahashi Y, Takayama T, et al. Transfection of glutathione S-transferase (GST)-pi anti sense complementary DNA increase sthesensitivity of a colon cancer cell line to adriamycin, cisplatin, melphalanandetoposide. Cancer Res. 1996;56:3577-3582.
  • Goto S, Ieda T, Cho S, Oka M, Kohno S, Kondo T. Over expression of glutathione S-transferase pi enhances the adduct formation of cisplatin with glutathione in human cancer cells. Free RadicRes. 1999;31:549-558.
  • Fengxi S, Xiaogu H, Weijuan J, Chang G, Erwei S, Peter H. Glutathione S Transferase π indicates chemotherapy resistance in breast cancer. Journal of Surgical Research. 2003;113:102-108.
  • Jingxiang H, Puay-Hoon T, Jayabaskar T, Boon-Huat B. Prognostic significance of glutathione s-transferase-pi in invasive breast cancer. Mod Pathol 2003;16(6):558–565.
  • Gilbert L, Elwood L, Merino M, et al. A pilot study of pi-class glutathione S-transferase expression in breastcancer: correlation with estrogen receptor expression and prognosis in node-negative breast cancer. JCO. 1993;11(1):49-58.
  • García-Closas M, Kelsey KT, Hankinson SE, et al. Glutathione S-Transferase Mu and Theta Polymorphisms and Breast Cancer Susceptibility. JNCI J Natl Cancer Inst. 1999;91(22):1960-1964.
  • Nakopoulou L, Alexandrou P, Stefanaki K, Panayotopoulou E, Lazaris AC, Davaris PS. Immunohistochemical expression of caspase-3 as an adverse indicator of the clinical outcome in human breast cancer. Patobiology. 2001;69:266-273.
  • Gorczyca W, Markiewski M, Kram A, Tuziak T, Domagala W. Immunohistochemic alanalysis of BCL-2 and p53 expression in breast carcinomas: their correlation with Ki-67 growth fraction. Virchows Archiv. 1995;426(3):229–233.
  • Pilco-Ferreto N, Calaf GM. Influence of doxorubicin on apoptosis and oxidative stress in breast cancer cell lines. Int J Oncol. 2016;49(2):753-62
  • Nooter K, Brutel G, Riviere de la M, et al. The prognostic significance of expression of the multidrug resistance-associated protein (MRP) in primary breast cancer. Br J Cancer. 1997;76(4):486–493.
  • Taniguchi K, Wada M, Kohno K, et al. A human canalicular multi specific organic anion transporter (cmoat) gene is overexpressed in cisplatin-resistant human cancer cell lines with decreased drug accumulation. Cancer Res. 1996;56:4124-9.
  • Kool M, de Haas M, Scheffer GL, et al. Analysis of Expression of cMOAT (MRP2), MRP3, MRP4 and MRP5 homologues of the multidrug resistance-associated protein gene (MRP1), in human cancer cell lines. Cancer Res. 1997;57:3537-47.
  • Ishikawa T, Ali Osman F. Glutathione-associatedcis-diammine dichloro platinum (II) metabolism and ATP-dependent efflux from leukemia cells. Molecular characterization of glutathione-platinum complex and its biological significance. The Journal of Biological Chemistry. 1993;268:20116-20125.
  • Faneyte IF, Kristel PM, van de Vijver MJ. Multidrug resistance associated genes MRP1, MRP2 and MRP3 in primary and anthracycline exposed breast cancer. Anticancer Research. 2004;24:2931-2940.
  • Leah C, Young B, Campling G, et al. Multi drug resistance proteins MRP3, MRP1 and MRP2 in lung cancer correlation of protein levels with drug response and messenger RNA levels. Clin Cancer Res. 2001;7:1798.
  • Larkin A, O'Driscoll L, Kennedy S, et al. Investigation Of MRP-1 Protein And MDR-1 P-Glycoprotein expression in invasive breast cancer: a prognostıc study. J. Cancer. 2004;112:286 –294.
  • Filipits M, Suchomel RW, Dekan G, et al. MRP and MDR1 Gene Expression in Primary Breast Carcinomas. Clin Cancer Res. 1996;2:1231-1237.
  • Keith WN, Stallard S, Brown R. Expression of mdr1 and gst-pi in human breast tumours: comparisonto invitro chemosensitivity. Br J Cancer. 1990;61(5):712–716.
  • Terrier P, Townsend AJ, Coindre JM, Triche TJ, Cowan KH. An Immunohistochemical study of pi class glutathione s-transferase expression in normal human tissue. American Journal of Pathology. 1990;137:4.
  • Juliano RL, Ling V. A surface glycoprotein modulation drug permeability in Chinese hamster ovary cell mutants. Biochim. Biophys. Acta. 1976;455(1):152-162.
  • Ueda K, Cardarelli C, Gottesman MM, Pastan I. Expression of and full-length c DNA fort he human MDR1 gene confers resistance tocolchicino, doxorubicin and vinblastine. Proc. Natl. Acad. Sci. USA. 1987;84:3004-3008.
  • Sun JH, Cheng ZG, Wang G, Hao SJ, Zou XHMJ. Multidrug resistance P-glycoprotein: crucial significance in drug disposition and interaction. Med. Sci. Monit. 2004;10:5-14.
  • Cole SPC, Bhardwaj G, Gerlach JH, et al. Over expression of a transporter gene in a multi drug-resistant human lung cancer cell line. Science. 1992;258:1650-1654

Investigation of GST Isoenzymes, Multi-Drug Resistance Proteins and Apoptotic Effect in MCF-7 Human Breast Cancer Cell Line Before and After Doxorubicin Treatment

Yıl 2020, Sayı: 10, 1 - 18, 17.04.2020
https://doi.org/10.38079/igusabder.631695

Öz

Aim: In the treatment of cancer, one of the problems encountered in clinical therapy of tumor cells had developed resistance against chemotherapy patients. The ABC (ATP-binding cassette) protein that helps drugs outside the cell membrane and has shown resistance of tumor cells to these drugs, is one of the most important members of transporter proteins. Besides that drug resistance protein is known to be effective on other intracellular proteins. In this context, the alkylating functionality at developing resistance to the cancer drug has been reported that increased level intracellular glutathione and glutathione S- conjugates have a role in cancer chemoresistance.
Method: In this study, the expression of GST isoenzyme family and ABC transporter proteins in drug-treated and untreated breast cancer MCF-7 cell line was examined by the immunocytochemical method.
Results: It was observed that the expression of GSTP1, GSTT1, GSTM1, GSTA1, GSTO1, GSTZ1 and GSTK1 protein in drug-treated breast cancer MCF-7 cell line was higher than that of untreated cells, no GSTS1 protein was found. The MRP 2,3,6,7 protein expressions in drug-treated breast cancer MCF-7 cell line was found to be higher than in the untreated cells, no MDR, MRP1 proteins were found. Also, bcl-2, p53, p38, caspase-3 protein expressions in drug-treated breast cancer MCF-7 cell line were found to be higher than in untreated cells.
Conclusion: MRP-2,3,6,7 of the ABC superfamily have a role in the development of breast cancer, and GST isoenzymes are important in drug resistance.

Proje Numarası

2015/125

Kaynakça

  • Tozkoparan B, Peri Aytaç S. Kanser kemoterapisinde terapötik hedef olarak glutatyon S-transferazlar. Hacettepe Üniversitesi, Eczacılık Fakültesi Dergisi. 2007;7(2):139-164.
  • Dünyada Kanser İstatistikleri. Türk Kanser Araştırma ve Savaş Kurumu (TKASKD).http://turkkanser.org.tr. Yayınlanma tarihi 1995. Erişim tarihi 20.09.2019.
  • Onur H. Tıbbi Onkoloji Kitabı. Ankara Üniversitesi Tıp Fakültesi: Antıp Yayınları; 2005.
  • Gate L, Tew KD. Glutathione S-transferases as emerging targets, Expert Opin Ther. Targets. 2001;5(4):477-489.
  • Szakács G, Paterson, JK, Ludwig JA, Booth-Genthe C, Gottesman MM. Targeting multidrug resistance in cancer. Nat Rev Drug Discov. 2006;5(3):219-34.
  • Stavrovskaya AA, Stromskaya TP. Transport proteins of the ABC familya and multidrug resistance of tumor cells. Biochemistry (Mosc). 2008;73(5):592-604.
  • Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics. CA Cancer J Clin 2009;59(4):225-249. doi: 10.3322/caac.20006.
  • Tuncer M. Significance of cancer in Turkey, the burden of disease and cancer control policies. Cancer Control in Turkey, Ankara, Onur Press, Health Ministry Publication. 2008:74;5-9.
  • Marie JP, Legrand O. Drug resistance in acute leukaemia and reversion. Turk J Med Sci. 2003;33:271-279.
  • Turgut S, Yaren A, Kursunluoğlu R, Turgut G. MDR1 C3435T polymorphism in patients with breast cancer. Arch Med Res. 2007:38;539-544.
  • Eaton DL, Bammler TK. Concise Review of glutathione s-transferase and their significance to toxicology. Toxicological Sciences. 1999;49:156-164.
  • Oguztuzun S, Abu-Hijleh A, Coban T, et al. GST isoenzymes in matched normal and neoplastic breast tissue. Neoplasma. 2011;58(4):304-310.
  • Buller Al, Clapper M, Smith T, Tew KD Glutathione S-trasnferase in nitrogen mustard-resistance and-sensitive. Molecular Pharmacology. 1987;31(6):575-578.
  • Oguri T, Fujimara Y, Katoh O, et al. Glutathione S-transferase gene expression and platinum drug exposure in human lung cancer. Cancer Letters. 2000;156:93-99.
  • Ban N, Takahashi Y, Takayama T, et al. Transfection of glutathione S-transferase (GST)-pi anti sense complementary DNA increase sthesensitivity of a colon cancer cell line to adriamycin, cisplatin, melphalanandetoposide. Cancer Res. 1996;56:3577-3582.
  • Goto S, Ieda T, Cho S, Oka M, Kohno S, Kondo T. Over expression of glutathione S-transferase pi enhances the adduct formation of cisplatin with glutathione in human cancer cells. Free RadicRes. 1999;31:549-558.
  • Fengxi S, Xiaogu H, Weijuan J, Chang G, Erwei S, Peter H. Glutathione S Transferase π indicates chemotherapy resistance in breast cancer. Journal of Surgical Research. 2003;113:102-108.
  • Jingxiang H, Puay-Hoon T, Jayabaskar T, Boon-Huat B. Prognostic significance of glutathione s-transferase-pi in invasive breast cancer. Mod Pathol 2003;16(6):558–565.
  • Gilbert L, Elwood L, Merino M, et al. A pilot study of pi-class glutathione S-transferase expression in breastcancer: correlation with estrogen receptor expression and prognosis in node-negative breast cancer. JCO. 1993;11(1):49-58.
  • García-Closas M, Kelsey KT, Hankinson SE, et al. Glutathione S-Transferase Mu and Theta Polymorphisms and Breast Cancer Susceptibility. JNCI J Natl Cancer Inst. 1999;91(22):1960-1964.
  • Nakopoulou L, Alexandrou P, Stefanaki K, Panayotopoulou E, Lazaris AC, Davaris PS. Immunohistochemical expression of caspase-3 as an adverse indicator of the clinical outcome in human breast cancer. Patobiology. 2001;69:266-273.
  • Gorczyca W, Markiewski M, Kram A, Tuziak T, Domagala W. Immunohistochemic alanalysis of BCL-2 and p53 expression in breast carcinomas: their correlation with Ki-67 growth fraction. Virchows Archiv. 1995;426(3):229–233.
  • Pilco-Ferreto N, Calaf GM. Influence of doxorubicin on apoptosis and oxidative stress in breast cancer cell lines. Int J Oncol. 2016;49(2):753-62
  • Nooter K, Brutel G, Riviere de la M, et al. The prognostic significance of expression of the multidrug resistance-associated protein (MRP) in primary breast cancer. Br J Cancer. 1997;76(4):486–493.
  • Taniguchi K, Wada M, Kohno K, et al. A human canalicular multi specific organic anion transporter (cmoat) gene is overexpressed in cisplatin-resistant human cancer cell lines with decreased drug accumulation. Cancer Res. 1996;56:4124-9.
  • Kool M, de Haas M, Scheffer GL, et al. Analysis of Expression of cMOAT (MRP2), MRP3, MRP4 and MRP5 homologues of the multidrug resistance-associated protein gene (MRP1), in human cancer cell lines. Cancer Res. 1997;57:3537-47.
  • Ishikawa T, Ali Osman F. Glutathione-associatedcis-diammine dichloro platinum (II) metabolism and ATP-dependent efflux from leukemia cells. Molecular characterization of glutathione-platinum complex and its biological significance. The Journal of Biological Chemistry. 1993;268:20116-20125.
  • Faneyte IF, Kristel PM, van de Vijver MJ. Multidrug resistance associated genes MRP1, MRP2 and MRP3 in primary and anthracycline exposed breast cancer. Anticancer Research. 2004;24:2931-2940.
  • Leah C, Young B, Campling G, et al. Multi drug resistance proteins MRP3, MRP1 and MRP2 in lung cancer correlation of protein levels with drug response and messenger RNA levels. Clin Cancer Res. 2001;7:1798.
  • Larkin A, O'Driscoll L, Kennedy S, et al. Investigation Of MRP-1 Protein And MDR-1 P-Glycoprotein expression in invasive breast cancer: a prognostıc study. J. Cancer. 2004;112:286 –294.
  • Filipits M, Suchomel RW, Dekan G, et al. MRP and MDR1 Gene Expression in Primary Breast Carcinomas. Clin Cancer Res. 1996;2:1231-1237.
  • Keith WN, Stallard S, Brown R. Expression of mdr1 and gst-pi in human breast tumours: comparisonto invitro chemosensitivity. Br J Cancer. 1990;61(5):712–716.
  • Terrier P, Townsend AJ, Coindre JM, Triche TJ, Cowan KH. An Immunohistochemical study of pi class glutathione s-transferase expression in normal human tissue. American Journal of Pathology. 1990;137:4.
  • Juliano RL, Ling V. A surface glycoprotein modulation drug permeability in Chinese hamster ovary cell mutants. Biochim. Biophys. Acta. 1976;455(1):152-162.
  • Ueda K, Cardarelli C, Gottesman MM, Pastan I. Expression of and full-length c DNA fort he human MDR1 gene confers resistance tocolchicino, doxorubicin and vinblastine. Proc. Natl. Acad. Sci. USA. 1987;84:3004-3008.
  • Sun JH, Cheng ZG, Wang G, Hao SJ, Zou XHMJ. Multidrug resistance P-glycoprotein: crucial significance in drug disposition and interaction. Med. Sci. Monit. 2004;10:5-14.
  • Cole SPC, Bhardwaj G, Gerlach JH, et al. Over expression of a transporter gene in a multi drug-resistant human lung cancer cell line. Science. 1992;258:1650-1654
Toplam 37 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Klinik Tıp Bilimleri
Bölüm Makaleler
Yazarlar

Arzu Kaya Koçdoğan 0000-0002-3689-3061

Serpil Oğuztüzün 0000-0002-5892-3735

Gülçin Güler Şimşek 0000-0001-7710-4631

Mustafa Türk 0000-0001-8202-090X

Proje Numarası 2015/125
Yayımlanma Tarihi 17 Nisan 2020
Kabul Tarihi 6 Ocak 2020
Yayımlandığı Sayı Yıl 2020 Sayı: 10

Kaynak Göster

JAMA Kaya Koçdoğan A, Oğuztüzün S, Güler Şimşek G, Türk M. MCF-7 İnsan Meme Kanseri Hücre Soyunda Doksorubisin Öncesi ve Sonrası GST İzozimlerinin, İlaç Dirençlilik Proteinlerinin ve Apoptotik Etkisinin Araştırılması. IGUSABDER. 2020;:1–18.

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