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

Abstract

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.

Keywords

• Breast cancer,GST,MDR,MRP,immunocytochemistry

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

Abstract

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. 

Keywords

• Meme Kanseri,GST,MDR,MRP,immunositokimya

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