Apoptosis Induction Through Increased TRPV1 Activation by Synergic Effect of Melatonin and Doxorubicin in Human Osteosarcoma and Chondrosarcoma Cell Lines

Yıl 2023, Cilt: 7 Sayı: 2, 145 - 152, 29.10.2023

Ahmet Koçak , Anıl Gülcü , İshak Suat Övey

https://doi.org/10.30565/medalanya.1313745

Öz

Aim: We aimed to reveal the role of doxorubicin (Dox), melatonin (Mel) and transient receptor potential Vanilloid 1 (TRPV1) channels in bone and cartilage cancer cells during the treatment process. Human Bone Osteosarcoma (Saos-2/An1) and Human Chondrosarcoma (Hs 819.T) cell lines were used to prepare in-vitro experiment models.

Methods: Both cell lines were cultured at 37°C. We have separated each cell line into five groups as follows: Controls, Dox, Dox+Capsazepine (Cpz), Dox+Melatonin (Mel), and combined Dox+Mel+Cpz given group. Capsaicin and capsazepine were added to cell culture mediums to activate or inactivate the TRPV1 channels, respectively. Cytosolic calcium, apoptosis, intracellular reactive oxygen, mitochondrial depolarization, caspase-3 and caspase-9 levels were measured.

Results: Increased apoptotic activity was detected in doxorubicin given cell lines (Group II) when compared with the controls (p˂0.001). There was also a significantly higher apoptotic level in Dox+Mel group (Group IV), when compared with only Dox given group (p˂0.001). TRPV1 inhibition applied groups (Group III and V) have had lower apoptotic levels than other drug administered groups (p˂0.001).

Conclusion: This study has indicated that apoptotic effects of Dox and Mel on both osteosarcoma and chondrosarcoma were strictly associated to TRPV1 channels, and that TRPV1 channels played an important role in whole mitochondria dependent pathways of apoptosis, which in turn may lead to increased intracellular Ca+2 levels and mitochondrial depolarization.

Anahtar Kelimeler

Osteosarcoma, chondrosarcoma, TRPV1, doxorubicin, melatonin

İnsan Osteosarkoma ve Kondrosarkoma Hücre Hatlarında Melatonin ve Doksorubisinin Sinerjik Etkisi Yoluyla Artan TRPV1 Etkinliği Üzerinden Apopitoz Uyarımı

Öz

Amaç: Kemik ve kıkırdak kanseri hücrelerinde doksorubisin (Dox) ve melatonin (Mel) ile birlikte geçici reseptör potansiyeli olan Vanilloid 1 (TRPV1) kanallarının tedavi sürecindeki rolünü ortaya çıkarmayı amaçladık. İn-vitro deney modellerinin hazırlanmasında İnsan Kemik Osteosarkomu (Saos-2/An1) ve İnsan Kondrosarkomu (Hs 819.T) hücre hatları kullanıldı.

Gereç ve Yöntem: Her iki hücre hattı da 37°C'de kültürlendi. Her hücre hattını aşağıdaki gibi beş gruba ayırdık: Kontroller, Dox, Dox+Kapsazepin (Cpz), Dox+Mel ve kombine Dox+Mel+Cpz verilen grup. Kapsaisin ve kapsazepin, sırasıyla TRPV1 kanallarını etkinleştirmek veya inaktive etmek için hücre kültürü ortamlarına eklendi. Sitozolik kalsiyum, apoptoz, hücre içi reaktif oksijen, mitokondriyal depolarizasyon, kaspaz 3 ve kaspaz 9 seviyeleri ölçüldü.

Bulgular: Doksorubisin (Dox) verilen hücre hatlarında (Grup II) kontrollere göre artmış apoptotik aktivite saptandı (p˂0.001). Ayrıca Dox+Mel grubunda (Grup IV), sadece Dox verilen grupla (p˂0.001) karşılaştırıldığında anlamlı derecede daha yüksek bir apoptotik seviye vardı. TRPV1 inhibisyonu uygulanan gruplar (Grup III ve V) diğer ilaç uygulanan gruplara göre daha düşük apoptotik düzeylere sahiptir (p˂0.001).

Sonuç: Bu çalışma, Dox ve Mel'in hem osteosarkom hem de kondrosarkom üzerindeki apoptotik etkilerinin TRPV1 kanalları ile kesin olarak ilişkili olduğunu ve TRPV1 kanallarının apoptozun tüm mitokondriye bağımlı yolaklarında önemli bir rol oynadığını ve bunun da hücre içi Ca+2 düzeylerinde artışa ve mitokondriyal depolarizasyona yol açabileceğini göstermiştir.

Anahtar Kelimeler

Osteosarkom, kondrosarkom, TRPV1, doksorubisin, melatonin

Kaynakça

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