Muscarinic Receptors as Targets for Metronomic Therapy in Ovarian Cancer

Abstract

Aim: In this study, the effects of muscarinic acetylcholine receptor (mAChR) agonist carbachol on the proliferation of cisplatin-resistant (A2780cis) and cisplatin-free (SKOV-3) ovarian cancer cell line were for the first time investigated to further evaluate the potential therapeutic effect of metronomic chemotherapy. Material and Methods: The inhibitory effect of carbachol on cell proliferation was detected using the xCELLigence Real-Time Cell Analyzer (RTCA) dual plate (DP) system. A preliminary study was conducted to determine the dose of carbachol 100 µM, cisplatin 1 µM, and two combination studies were carried out with 100 µM carbachol + cisplatin 1 µM and 100 µM carbachol + 10 µM atropine, over cancer cells without drugs was used as the control group. The cell proliferation curve was monitored for 96 hours. The cell index value of inhibition in cell proliferation was automatically measured every hour for each well using RTCA 1.2.1 software. Results: Co-administration of carbachol with cisplatin caused a decrease in cell number in both A2780cis and SKOV-3 cell lines in a time-dependent manner (p<0.001). Substantial cell death was observed in both cisplatin-resistant (A2780cis) and cisplatin-free (SKOV-3) cell lines within 24 hours after carbachol with cisplatin application and this continued at the 96th hour. Conclusion: The findings of this study confirm the notion that mAChRs can be considered as therapeutic targets for metronomic therapy in ovarian cancer, as well as the usefulness of a muscarinic agonist as a repositioning drug in the treatment of such tumors.

Keywords

• Ovary cancer;
• mAChR;
• Carbachol;
• SKOV-3;
• A2780CIS;
• cisplatin

Yumurtalık Kanserinde Metronomik Tedavi Hedefi Olarak Muskarinik Reseptörler

Abstract

Amaç: Bu çalışmada, muskarinik asetilkolin reseptörü (mAChR) agonisti karbakolun, sisplatine dirençli (A2780cis) ve sisplatine dirençsiz (SKOV-3) yumurtalık kanseri hücre hatlarının proliferasyonu üzerindeki etkileri, metronomik kemoterapinin potansiyel terapötik etkisini daha ileri düzeyde değerlendirmek üzere ilk kez araştırıldı. Gereç ve Yöntemler: Karbakolün hücre proliferasyonu üzerindeki inhibitör etkisi, xCELLigence Real-Time Cell Analyzer (RTCA) dual plate (DP) sistemi kullanılarak tespit edildi. 100 µM karbakol ve 1 µM sisplatin dozunu belirlemek için bir ön çalışma yapıldı ve 100 µM karbakol + 1 µM sisplatin ve 100 µM karbakol + 10 µM atropin olmak üzere iki kombinasyon çalışması yapıldı, ilaçsız yumurtalık kanser hücreleri ise kontrol grubu olarak kullanıldı. Hücre proliferasyon eğrisi 96 saat izlendi. Hücre proliferasyonundaki inhibisyonun hücre indeksi değeri, RTCA 1.2.1 yazılımı ile her kuyu için her saat otomatik olarak ölçüldü. Bulgular: Karbakolun sisplatin ile birlikte uygulanması hem A2780cis hem de SKOV-3 hücre hatlarında zamana bağlı olarak hücre sayısında azalmaya neden olmuştur (p<0.001). Sisplatin ile karbakol uygulamasından sonraki 24 saat içinde sisplatine dirençli (A2780cis) ve sisplatine dirençsiz (SKOV-3) her iki hücre hattında da önemli düzeyde hücre ölümü gözlenmiş ve bu durum 96. saatte de devam etmiştir (p<0.001). Sonuç: Bu çalışmanın bulguları, mAChR'lerin yumurtalık kanserinde metronomik tedavi için terapötik hedefler olarak kabul edilebileceği fikrini ve ayrıca bu tür tümörlerin tedavisinde bir muskarinik agonistin bir yeniden konumlandırma ilacı olarak kullanışlılığını doğrulamaktadır.

Keywords

• Yumurtalık kanseri;
• mAChR;
• Karbakol;
• SKOV-3;
• A2780CIS;
• Sisplatin

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