İnsan Meme Kanseri Hücre Hatlarında Verapamil’in Dosetaksel, Gemsitabin ve Karboplatin ile Kombine Tedavisinin, Sitotoksik ve Apopitotik Etkilerinin Araştırılması

Yıl 2024, Cilt: 50 Sayı: 3, 509 - 518, 12.01.2025

Dilek Yeğin , Engin Ulukaya

https://doi.org/10.32708/uutfd.1549517

Öz

Kemoterapi ilaçlarına karşı direnç, kanser tedavisindeki en yaygın ve ciddi klinik sorundur. Başarılı kanser tedavisindeki en büyük zorluk olan bu olguya çoklu ilaç direnci denir. Çoklu ilaç direncinin yaygın bir nedeni P-glikoproteinin aşırı ekspresyonudur. Verapamil, P-glikoprotein ekspresyonunu azaltan bir kalsiyum kanal blokeridir. Bu çalışma, Verapamilin çeşitli dozlarda, MDA-MB-231 ve MCF-7 meme kanseri hücre hatlarını kullanarak, kemoterapötiklerin sitotoksik potansiyelini arttırma etkisini araştırdı. İnsan meme kanseri hücre hatları MDA-MB-231 ve MCF-7, Dosetaksel, Gemsitabin ve Karboplatin kombinasyonlarıyla tedavi edildi. Hücre canlılığı MTT yöntemi kullanılarak değerlendirildi. Apopitozun varlığı (M30 Antijeni) ELISA yöntemi kullanılarak tespit edildi. Apopitoz tespiti için Annexin-V floresan konjugatları kullanıldı. Primer nekroz veya geç apopitoz (sekonder nekroz) nedeniyle hasarlı membranlara sahip hücrelerin DNA'sına propidyum iyodür, sekonder boya olarak kullanıldı ve floresan mikroskopisi ile analiz edildi. Çalışmada Verapamil'in hem MDA-MB-231 hem de MCF-7 hücre hatlarında Dosetaksel ve Karboplatinin sitotoksik etkilerini artırdığı tespit edildi. Ek olarak Verapamil, hücre tipine bağlı olarak Gemsitabinin etkinliğini değiştirdi. Verapamil, MCF-7 hücre hattında kaspazla parçalanan sitokeratin 18'i arttırdı. MCF-7 hücrelerinde Gemsitabin-Verapamil kombinasyonunun kullanılmasının sitotoksik etkiyi artırma açısından etkili bir strateji olabileceği düşünüldü. Ancak bu sonuçların daha fazla ve farklı deneylerle doğrulanması gerekmektedir.

Anahtar Kelimeler

Verapamil, Çoklu İlaç Direnci, Meme Kanseri, P-Glikoprotein, Kemoterapi, Hücre Kültürü

Investigation of Cytotoxic and Apoptotic Effects of Combined Treatment of Verapamil with Docetaxel, Gemcitabine and Carboplatin in Human Breast Cancer Cell Lines

Öz

Resistance to chemotherapeutic drugs is the most common and serious clinical problem in cancer treatment. This phenomenon, which is the major challenge in successful cancer therapy, is referred to as multidrug resistance. A frequent reason for multidrug resistance is the heightened production of P-glycoprotein. Verapamil, a calcium channel antagonist, diminishes P-glycoprotein levels. This study investigated the effect of Verapamil at various doses on enhancing the cytotoxic potential of chemotherapeutics using MDA-MB-231 and MCF-7 breast cancer cell lines. Human breast cancer cell lines MDA-MB-231 and MCF-7 were treated with combinations of Docetaxel, Gemcitabine, and Carboplatin. Cell viability was assessed using the MTT method. The presence of apoptosis (M30 Antigen) was detected using the ELISA method. Annexin-V fluorescent conjugates were used for apoptosis detection. Propidium iodide, which binds to the DNA of cells with damaged membranes due to primary necrosis or late apoptosis (secondary necrosis), was used as a secondary dye and analyzed by fluorescence microscopy. In this study, Verapamil was observed to amplify the cytotoxicity of Docetaxel and Carboplatin in both MDA-MB-231 and MCF-7 cell lines. Furthermore, Verapamil influenced the effectiveness of Gemcitabine in a cell type-dependent manner. Verapamil increased caspase-cleaved cytokeratin 18 in MCF-7 cell line. It is thought that the use of Gemcitabine-Verapamil combination in MCF-7 cells may be an effective strategy in terms of increasing the cytotoxic effect. However, these results need to be confirmed with more and different experiments.

Anahtar Kelimeler

Verapamil, Multidrug Resistance, Breast Cancer, P-Glycoprotein, Chemotherapy

Kaynakça

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