Özay GÜLEŞ, Esra BİLİCİ, Emira KURBASEVIC, Ömer Faruk LENGER, Murat BOYACIOĞLU, Erkmen Tuğrul EPİKMEN

G-361 insan melanom kanseri hücre hattında TRPV4 antagonisti RN-1734'ün sitotoksik aktivitesi

Amaç: Hücre içi kalsiyum (Ca2+) sinyali, hücre çoğalması ve farklılaşması, gen transkripsiyonu, oksidatif stres, antioksidan sistem ve apoptoz gibi birçok hücresel olayda rol oynar. Geçici reseptör potansiyeli vanilloid 4 (TRPV4) kanalları, seçici olmayan bir katyon (Ca2+) kanalıdır. Bu çalışmada, TRPV4 antagonisti olan RN-1734'ün G361 insan melanom kanser hücre hattındaki sitotoksik aktivitesinin araştırılması amaçlandı. Gereç ve Yöntem: RN-1734'ün 1, 5, 25, 50 ve 100 uM konsantrasyonlarında G361 hücre canlılığı üzerindeki etkisi 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolyum bromid (MTT) yöntemi ile ölçüldü. Total antioksidan kapasite (TAS) ve total oksidan kapasite (TOS) seviyeleri hazır ticari bir kit kullanılarak belirlendi ve sonrasında oksidatif stres indeksi (OSI) değeri hesaplandı. RN-1734'ün apoptotik etkilerini tespit etmek için Bcl-2, Bax ve p53 ekspresyon seviyeleri ile kaspaz 3 ve -8 aktiviteleri kantitatif real-time PCR yöntemi ile belirlendi. Bulgular: G361 hücre canlılığı, kontrol grubuna (%100.00) kıyasla tüm RN-1734 gruplarında (1, 5, 25, 50 ve 100 μM) önemli ölçüde %82.72, 72.81, 56.36, 39.16 ve 18.96'ya düştü. Yine IC50 (39.48 μM), konsantrasyonunda RN-1734 uygulaması (3.35 mmol/g prot.-TAS, 45.87 μmol/g prot.-TOS, 1501.97 AU-OSI) kontrol grubuna gore TAS'I (2.17 mmol/g prot.) artırmış, TOS (55.41 μmol/g prot.) ve OSI'yi (3142.76 AU) azaltmıştır. Sonuç: Bulgularımız, RN-1734'ün, G361 insan melanom kanser hücrelerinin canlılığını azaltarak, melanom için yeni bir terapötik yaklaşım olabileceğini göstermektedir.

Anahtar Kelimeler:

Hücre canlılığı, G361 insan melanom kanser hücreleri, oksidatif stres, RN-1734, TRPV4

Cytotoxic activity of TRPV4 antagonist RN-1734 in G-361 human melanoma cancer cell line

Purpose: Intracellular calcium (Ca2+) signaling plays a role in many cellular events, such as cell proliferation and differentiation, gene transcription, oxidative stress, the antioxidant system, and apoptosis. Transient receptor potential vanilloid 4 (TRPV4) channels are non-selective cation (Ca2+) channels. The present study aims to investigate the cytotoxic activity of RN-1734, a transient receptor potential vanilloid 4 (TRPV4) antagonist, in the G361 human melanoma cancer cell line. Materials and Methods: The effects of RN-1734 on G361 cell viability at concentrations of 1, 5, 25, 50, and 100 μM were measured using the 3-(4,5-dimethylthiazol-2-il)-2,5-diphenyltetrazolium bromide (MTT) method. Total antioxidant status (TAS) and total oxidant status (TOS) levels were determined using a ready-made commercial kit, after which oxidative stress index (OSI) values were calculated. To determine the apoptotic effects of RN-1734, Bcl-2, Bax, and p53 expression levels, caspase-3 and -8 activities were examined via quantitative real-time PCR analysis. Results: G361 cell viability significantly decreased to 82.72, 72.81, 56.36, 39.16 and 18.96% in RN-1734 groups (1, 5, 25, 50 and 100 μM) compared to the control group (100.00%). At IC50 concentration (39.48 μM), RN-1734 application (3.35 mmol/g prot.-TAS, 45.87 μmol/g prot.-TOS, and 1501.97 AU-OSI) increased the TAS level (2.17 mmol/g prot.) and decreased the TOS level (55.41 μmol/g prot.) and OSI value (3142.76 AU) compared to the control group. Conclusion: Our findings show that RN-1734 may be a novel therapeutic approach to treating melanoma by decreasing the cell viability of G361 human melanoma cancer cells.

Keywords:

Cell viability, G361 human melanoma cancer cells, oxidative stress, RN-1734, TRPV4,

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