Determination of mechanisms of multiple drug resistance in MCF-7 cell line developed against paclitaxel and vincristine by microarray analysis

Abstract

OBJECTIVES Acquired or intrinsic multiple drug resistance (MDR) is one of the limitations for successful chemotherapy. In this study, MDR mechanisms developed after paclitaxel and vincristine application in MCF-7 cell line were investigated. METHODS Paclitaxel and vincristine were applied to MCF-7 cell line in stepwise manner. The resistance levels of the cells were determined by XTT test. The levels of all of the genes in resistant and sensitive cells were determined by microarray assay. RESULTS MCF-7/400nMPac and MCF-7/120nMVinc cells are 150- and 30-fold resistant compared to sensitive MCF-7. Upregulation of MDR1 gene is found to be important in resistance. The expression levels of the genes encoding detoxifying enzymes were upregulated and some apoptotic gene levels were downregulated. Changes in expression levels of metastatic genes, some oncogenes, cell cycle, and tubulin-related genes were found. CONCLUSION According to these results, change in expression level of MDR1 gene seems to be important in resistance in MCF-7 cells.

Keywords

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Meme kanseri MCF-7 hücre hattında paklitaksel ve vinkristin'e karşı gelişmiş çoklu ilaç direnci mekanizmalarının mikrodizin analizi ile belirlenmesi

Öz

AMAÇ Sonradan kazanılan veya tedavi öncesi varolan çoklu ilaç dirençliliği (ÇİD) kemoterapide başarıyı engelleyen nedenlerden biridir. Bu çalışmada, paklitaksel ve vinkristin'e karşı gelişen ÇİD mekanizmaları MCF-7 hücre hattında araştırıldı. GEREÇ VE YÖNTEM MCF-7 hücreleri önce kademeli olarak paklitaksel ve vinkristin doz artışlarına maruz bırakıldı. Hücrelerin direnç seviyeleri XTT testi ile belirlendi. Tüm dirençli ve duyarlı hücrelerde genlerin düzeyleri mikrodizin yöntemiyle belirlendi. BULGULAR MCF-7/400nMPak ve MCF-7/120nMVink hücre hatlarının orjinal hücrelere göre 150 ve 30 kat daha dirençli oldukları bulundu. MDR1 genin ifadesindeki artışın dirençlilik mekanizmaları arasında önemli olduğu bulundu. Hücrelerde toksik maddelerin metabolize edilmesiyle ilgili genlerin ifade düzeylerinde artışlar gözlenirken, apoptozla ilgili bazı genlerin ifadelerinde azalmalar olduğu görüldü. Metastaz genlerinde, bazı onkojenlerde, hücre döngüsünü düzenleyen ve tübülin metabolizmasıyla ilgili genin ifade düzeylerinde değişimler saptandı. SONUÇ ÇİD'de MDR1 genindeki değişikliğin MCF-7'de paklitaksel ve vinkristin dirençliliğinde temel mekanizma olduğu görülmüştür.

Anahtar Kelimeler

• Apoptotik yolak, beta tübülin, çoklu ilaç dirençliliği, MCF-7, mikrodizin analizi

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