Adhesion of beta 1 integrin to fibronectin regulates CAM-DR phenotype via p21 in HL60 acute myeloid leukemia (AML) cells

Genel Bilgiler: İlaç direnci, kanser tedavisinin başarısında büyük zorluktur. Hücre adezyon aracılı ilaç direnci (CAM-DR), kanser hücrelerinin mikroçevre ile ilişkisiyle gelişen, çeşitli tümörlerde gösterilen yeni bir ilaç direncidir. Bu çalışma da, HL60 AML hücrelerinde CAM-DR fenotipi analiz edildi. Yöntem: HL60 hücrelerin fibronektine bağlanması kolorimetrik adezyon deneyi ile yapıldı. Doksorubisin ile indüklenen apoptozis ve hücre siklusu akım sitometrik analiz ile belirlendi. [3H]-timidin inkorporasyon deneyi ile proliferasyon hızı değerlendirildi. Hücre siklusunu kontrol eden proteinler, western blot ve RT-PCR analizi ile belirlendi. Bulgular: HL60 hücrelerinin a4pM and oc5pi integrin aracılığı ile fibronektine bağlanmasıyla CAM-DR fenotipi oluşmuştur, aderent HL60 hücreleri doksorubisin ile tetiklenen apoptozise dirençli hale geçmiştir. Fibronektine aderan/yapışan HL60 hücrelerinde hücre siklusunun GQ/G, fazında birikimi ile hücre proliferasyonu durmuştur. Buna karşılık, fibronektinden" sökülen hücreler 8 saat sonra tekrar sentez fazına girerek apoptozise duyarlı hale gelmiştir. Hücre siklusunun GQ/G, kontrol noktalarının analizi, CAM-DR fenotipinin p21waf/cıp proteini ile düzenlendiğini göstermiştir. Sonuç: Bu çalışmada, CAM-DR fenotipininin AML blastlarında apoptozisi azaltan ve proliferasyonu durduran geri dönüşümlü bir ilaç direnci mekanizması olabileceğini gösterdik.

Beta 1 integrinin fibronektine adezyonu HL60 akut myeloid lösemi (AML) hücrelerinde p21aracılığı ile CAM-DR fenotipini düzenler

Aims: Drug resistance is a major obstacle for a successful cancer therapy. Cell adhesion mediated drug resistance (CAM-DR) is a novel type of drug resistance and generated via interaction of cancer cells with the microenvironment. In this study, CAM-DR phenotype was analyzed in HL60 acute myeloid leukemia (AML) cells. Materials and Methods: Fibronectin (FN) adherence of HL60 cells was tested by a colorimetric adhesion assay. Flow cytometry analyses were performed to evaluate doxorubicin-induced apoptosis and to determine cell cycle status. Proliferation rate was evaluated by [3H]-thymidine incorporation assay. Western blot and RTPCR were used for analysis of the factors involved in cell cycle control. Results: Binding of HL60 to FN via a4pM and a5|31 integrins exerted a CAM-DR phenotype, which shows resistance to apoptosis triggered by doxorubicin. FN-adherent HL60 cells accumulated in the GQ/G, phase of cell cycle and stopped proliferation. However, after detachment from FN, cells entered S phase, proliferated, and became sensitive to apoptosis. The analysis of the factors involved in the Gg/G, cell cycle checkpoint showed that CAM-DR phenotype might be regulated mainly by p21waf/cip. Conclusions: Here we showed that CAM-DR may also represent a reversible drug resistance mechanism that decreases apoptosis and causes growth arrest in AML blasts.

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