Öz
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
Kaynakça
- Kim GH, Fang XQ, Lim WJ, Park J, Kang TB, Kim JH et al. Cinobufagin suppresses melanoma cell growth by inhibiting LEF1. Int J Mol Sci. 2020;21:6706.
- Siegel R, DeSantis C, Virgo K, Stein K, Mariotto A, Smith T et al. Cancer treatment and survivorship statistics. CA Cancer J Clin. 2012;62:220-41.
- Park KH, Choi JH, Song YS, Kim GC, Hong JW. Ethanol extract of asiasari radix preferentially induces apoptosis in G361 human melanoma cells by differential regulation of p53. BMC Complement Altern Med. 2019;19:231.
- Leonardi GC, Falzone L, Salemi R, Zanghì A, Spandidos DA, Mccubrey JA et al. Cutaneous melanoma: From pathogenesis to therapy. Int J Oncol. 2018;52:1071-80.
- Parkin DM, Mesher D, Sasieni P. Cancers attributable to solar (ultraviolet) radiation exposure in the UK in 2010. Br J Cancer. 2011;105:66-9.
- Zhang W, Xie X, Huang Z, Zhong X, Liu Y, Cheong KL et al. The integration of single-cell sequencing, TCGA, and GEO data analysis revealed that PRRT3-AS1 is a biomarker and therapeutic target of SKCM. Front Immunol. 2022;13:919145.
- Chen YN. Dacarbazine inhibits proliferation of melanoma FEMX-1 cells by up-regulating expression of miRNA-200. Eur Rev Med Pharmacol Sci. 2017;21:1191-7.
- Yanarojana M, Nararatwanchai T, Thairat S, Tancharoen S. Antiproliferative activity and induction of apoptosis in human melanoma cells by Houttuynia cordata thunb extract. Anticancer Res. 2017;37:6619-28.
- Stewart TA, Yapa KT, Monteith GR. Altered calcium signaling in cancer cells. Biochim Biophys Acta. 2015;1848:2502-11.
- Yu S, Huang S, Ding Y, Wang W, Wang A, Lu Y. Transient receptor potential ion-channel subfamily V member 4: a potential target for cancer treatment. Cell Death Dis. 2019;10:497.
- Thoppil RJ, Adapala RK, Cappelli HC, Kondeti V, Dudley AC, Meszaros JG et al. TRPV4 channel activation selectively inhibits tumor endothelial cell proliferation. Sci Rep. 2015;5:14257.
- Peters AA, Jamaludin SYN, Yapa K, Chalmers S, Wiegmans AP, Lim HF et al. Oncosis and apoptosis induction by activation of an overexpressed ion channel in breast cancer cells. Oncogene 2017;36:6490-500.
- Fang Y, Liu G, Xie C, Qian K, Lei X, Liu Q et al. Pharmacological inhibition of TRPV4 channel suppresses malignant biological behavior of hepatocellular carcinoma via modulation of ERK signaling pathway. Biomed Pharmacother. 2018;101:910-9.
- Çakır M, Erden Y. MCF-7 ve PC-3 hücre hatlarında, hücre canlılığı ve DNA hasarı üzerine TRPV4 antagonisti RN1734’ün etkileri. Bozok Med J. 2019;9:134-9.
- Xu X, Lai Y, Hua ZC. Apoptosis and apoptotic body: disease message and therapeutic target potentials. Biosci Rep. 2019;39:BSR20180992.
- Obeng E. Apoptosis (programmed cell death) and its signals – A review. Braz J Biol. 2021;81:1133-43.
- Kashyap D, Garg VK, Goel N. Intrinsic and extrinsic pathways of apoptosis: Role in cancer development and prognosis. Adv Protein Chem Struct Biol. 2021;125:73-120.
- Reed JC. Bcl-2 family proteins: regulators of apoptosis and chemoresistance in haematologic malignancies. Semin Haematol. 1997;34:9-19.
- Sharma A, Boise LH, Shanmugam M. Cancer metabolism and the evasion of apoptotic cell death. Cancers (Basel). 2019;11:1144.
- Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55-63.
- Koran K, Tekin Ç, Çalışkan E, Tekin S, Sandal S, Görgülü AO. Synthesis, structural and thermal characterizations and in vitro cytotoxic activities of new cyclotriphosphazene derivatives. Phosphorus Sulfur Silicon Relat Elem. 2017;192:1002-11.
- Borenfreund E, Puerner JA. A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NR-90). J Tissue Cult Methods. 1985;9:7-9.
- Kar F, Hacioglu C, Kacar S, Sahinturk V, Kanbak G. Betaine suppresses cell proliferation by increasing oxidative stress-mediated apoptosis and inflammation in DU-145 human prostate cancer cell line. Cell Stress Chaperones. 2019;24:871-81.
- Ulasli SS, Celik S, Gunay E, Ozdemir M, Hazman O, Ozyurek A et al. Anticancer effects of thymoquinone, caffeic acid phenethyl ester and resveratrol on A549 non-small cell lung cancer cells exposed to benzo(a)pyrene. Asian Pac J Cancer Prev. 2013;14:6159-64.
- White P, Kaestner PW. Gene expression analysis in diabetes resarch. Methods Mol Biol. 2009;560:239-61.
- Conover WJ. Practical Nonparametric Statistics. New York, Wiley, 1980.
- Patergnani S, Danese A, Bouhamida E, Aguiari G, Previati M, Pinton P et al. Various aspects of calcium signaling in the regulation of apoptosis, autophagy, cell proliferation, and cancer. Int J Mol Sci. 2020;21:8323.
- Liu X, Zhang P, Xie C, Sham KWY, Ng SSM, Chen Y et al. Activation of PTEN by inhibition of TRPV4 suppresses colon cancer development. Cell Death Dis. 2019;10:460.
- Erdoğan MA, Erbaş O. İyon kanalları ve kanser. FNG Bilim Tıp Dergisi 2018;4:49-62.
- Monteith GR, Prevarskaya N, Roberts-Thomson SJ. The calcium-cancer signalling nexus. Nat Rev Cancer. 2017;17:367-80.
- Bahari NN, Jamaludin SYN, Jahidin AH, Zahary MN, Mohd Hilmi AB. Assessment of TRPV4 channel and its role in colorectal canser cells. Biomed Pharmacol J. 2019;12:629-38.
- Wasilewski A, Krajewska U, Owczarek K, Lewandowska U, Fichna J. Fatty acid amide hydrolase (FAAH) inhibitor PF-3845 reduces viability, migration and invasiveness of human colon adenocarcinoma Colo-205 cell line: an in vitro study. Acta Biochim Pol. 2017;64:519-25.
- Fang Y, Liu G, Xie C, Qian K, Lei X, Liu Q et al. Pharmacological inhibition of TRPV4 channel suppresses malignant biological behavior of hepatocellular carcinoma via modulation of ERK signaling pathway. Biomed Pharmacother. 2018;101:910-19.
- Johansen JS, Harris AK, Rychly DJ, Ergul A. Oxidative stress and the use of antioxidants in diabetes: linking basic science to clinical practice. Cardiovasc Diabetol. 2005;4:5.
- Yilmazer A. Cancer cell lines involving cancer stem cell populations respond to oxidative stress. Biotechnol Rep. 2017;17:24-30.
- Sander CS, Hamm F, Elsner P, Thiele JJ. Oxidative stress in malignant melanoma and non-melanoma skin cancer. Br J Dermatol. 2003;148:913-22.
- Villalta PC, Rocic P, Townsley MI. Role of MMP2 and MMP9 in TRPV4-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 2014;307:L652-9.
- Hong Z, Tian Y, Yuan Y, Qi M, Li Y, Du Y et al. Enhanced oxidative stress is responsible for TRPV4-induced neurotoxicity. Front Cell Neurosci. 2016;232.
- Özşimşek A, Nazıroğlu M. The involvement of TRPV4 on the hypoxia-induced oxidative neurotoxicity and apoptosis in a neuronal cell line: Protective role of melatonin. Neurotoxicology. 2021;87:136-48.
- Wu Q, Lu K, Zhao Z, Wang B, Liu H, Zhang S et al. Blockade of transient receptor potential vanilloid 4 enhances antioxidation after myocardial ischemia/reperfusion. Oxid Med Cell Longev. 2019;7283683.
- Nazıroğlu M. A novel antagonist of TRPM2 and TRPV4 channels: Carvacrol. Metab Brain Dis. 2022;37:711–28
- Liu X, Zhang P, Xie C, Sham KWY, Ng SSM, Chen Y et al. Activation of PTEN by inhibition of TRPV4 suppresses colon cancer development. Cell Death Dis. 2019;10:460
Öz
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.
Anahtar Kelimeler
Cell viability G361 human melanoma cancer cells oxidative stress RN-1734 TRPV4
Teşekkür
We appreciate Prof. Sefa Çelik for supplying the lab equipment.
Kaynakça
- Kim GH, Fang XQ, Lim WJ, Park J, Kang TB, Kim JH et al. Cinobufagin suppresses melanoma cell growth by inhibiting LEF1. Int J Mol Sci. 2020;21:6706.
- Siegel R, DeSantis C, Virgo K, Stein K, Mariotto A, Smith T et al. Cancer treatment and survivorship statistics. CA Cancer J Clin. 2012;62:220-41.
- Park KH, Choi JH, Song YS, Kim GC, Hong JW. Ethanol extract of asiasari radix preferentially induces apoptosis in G361 human melanoma cells by differential regulation of p53. BMC Complement Altern Med. 2019;19:231.
- Leonardi GC, Falzone L, Salemi R, Zanghì A, Spandidos DA, Mccubrey JA et al. Cutaneous melanoma: From pathogenesis to therapy. Int J Oncol. 2018;52:1071-80.
- Parkin DM, Mesher D, Sasieni P. Cancers attributable to solar (ultraviolet) radiation exposure in the UK in 2010. Br J Cancer. 2011;105:66-9.
- Zhang W, Xie X, Huang Z, Zhong X, Liu Y, Cheong KL et al. The integration of single-cell sequencing, TCGA, and GEO data analysis revealed that PRRT3-AS1 is a biomarker and therapeutic target of SKCM. Front Immunol. 2022;13:919145.
- Chen YN. Dacarbazine inhibits proliferation of melanoma FEMX-1 cells by up-regulating expression of miRNA-200. Eur Rev Med Pharmacol Sci. 2017;21:1191-7.
- Yanarojana M, Nararatwanchai T, Thairat S, Tancharoen S. Antiproliferative activity and induction of apoptosis in human melanoma cells by Houttuynia cordata thunb extract. Anticancer Res. 2017;37:6619-28.
- Stewart TA, Yapa KT, Monteith GR. Altered calcium signaling in cancer cells. Biochim Biophys Acta. 2015;1848:2502-11.
- Yu S, Huang S, Ding Y, Wang W, Wang A, Lu Y. Transient receptor potential ion-channel subfamily V member 4: a potential target for cancer treatment. Cell Death Dis. 2019;10:497.
- Thoppil RJ, Adapala RK, Cappelli HC, Kondeti V, Dudley AC, Meszaros JG et al. TRPV4 channel activation selectively inhibits tumor endothelial cell proliferation. Sci Rep. 2015;5:14257.
- Peters AA, Jamaludin SYN, Yapa K, Chalmers S, Wiegmans AP, Lim HF et al. Oncosis and apoptosis induction by activation of an overexpressed ion channel in breast cancer cells. Oncogene 2017;36:6490-500.
- Fang Y, Liu G, Xie C, Qian K, Lei X, Liu Q et al. Pharmacological inhibition of TRPV4 channel suppresses malignant biological behavior of hepatocellular carcinoma via modulation of ERK signaling pathway. Biomed Pharmacother. 2018;101:910-9.
- Çakır M, Erden Y. MCF-7 ve PC-3 hücre hatlarında, hücre canlılığı ve DNA hasarı üzerine TRPV4 antagonisti RN1734’ün etkileri. Bozok Med J. 2019;9:134-9.
- Xu X, Lai Y, Hua ZC. Apoptosis and apoptotic body: disease message and therapeutic target potentials. Biosci Rep. 2019;39:BSR20180992.
- Obeng E. Apoptosis (programmed cell death) and its signals – A review. Braz J Biol. 2021;81:1133-43.
- Kashyap D, Garg VK, Goel N. Intrinsic and extrinsic pathways of apoptosis: Role in cancer development and prognosis. Adv Protein Chem Struct Biol. 2021;125:73-120.
- Reed JC. Bcl-2 family proteins: regulators of apoptosis and chemoresistance in haematologic malignancies. Semin Haematol. 1997;34:9-19.
- Sharma A, Boise LH, Shanmugam M. Cancer metabolism and the evasion of apoptotic cell death. Cancers (Basel). 2019;11:1144.
- Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55-63.
- Koran K, Tekin Ç, Çalışkan E, Tekin S, Sandal S, Görgülü AO. Synthesis, structural and thermal characterizations and in vitro cytotoxic activities of new cyclotriphosphazene derivatives. Phosphorus Sulfur Silicon Relat Elem. 2017;192:1002-11.
- Borenfreund E, Puerner JA. A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NR-90). J Tissue Cult Methods. 1985;9:7-9.
- Kar F, Hacioglu C, Kacar S, Sahinturk V, Kanbak G. Betaine suppresses cell proliferation by increasing oxidative stress-mediated apoptosis and inflammation in DU-145 human prostate cancer cell line. Cell Stress Chaperones. 2019;24:871-81.
- Ulasli SS, Celik S, Gunay E, Ozdemir M, Hazman O, Ozyurek A et al. Anticancer effects of thymoquinone, caffeic acid phenethyl ester and resveratrol on A549 non-small cell lung cancer cells exposed to benzo(a)pyrene. Asian Pac J Cancer Prev. 2013;14:6159-64.
- White P, Kaestner PW. Gene expression analysis in diabetes resarch. Methods Mol Biol. 2009;560:239-61.
- Conover WJ. Practical Nonparametric Statistics. New York, Wiley, 1980.
- Patergnani S, Danese A, Bouhamida E, Aguiari G, Previati M, Pinton P et al. Various aspects of calcium signaling in the regulation of apoptosis, autophagy, cell proliferation, and cancer. Int J Mol Sci. 2020;21:8323.
- Liu X, Zhang P, Xie C, Sham KWY, Ng SSM, Chen Y et al. Activation of PTEN by inhibition of TRPV4 suppresses colon cancer development. Cell Death Dis. 2019;10:460.
- Erdoğan MA, Erbaş O. İyon kanalları ve kanser. FNG Bilim Tıp Dergisi 2018;4:49-62.
- Monteith GR, Prevarskaya N, Roberts-Thomson SJ. The calcium-cancer signalling nexus. Nat Rev Cancer. 2017;17:367-80.
- Bahari NN, Jamaludin SYN, Jahidin AH, Zahary MN, Mohd Hilmi AB. Assessment of TRPV4 channel and its role in colorectal canser cells. Biomed Pharmacol J. 2019;12:629-38.
- Wasilewski A, Krajewska U, Owczarek K, Lewandowska U, Fichna J. Fatty acid amide hydrolase (FAAH) inhibitor PF-3845 reduces viability, migration and invasiveness of human colon adenocarcinoma Colo-205 cell line: an in vitro study. Acta Biochim Pol. 2017;64:519-25.
- Fang Y, Liu G, Xie C, Qian K, Lei X, Liu Q et al. Pharmacological inhibition of TRPV4 channel suppresses malignant biological behavior of hepatocellular carcinoma via modulation of ERK signaling pathway. Biomed Pharmacother. 2018;101:910-19.
- Johansen JS, Harris AK, Rychly DJ, Ergul A. Oxidative stress and the use of antioxidants in diabetes: linking basic science to clinical practice. Cardiovasc Diabetol. 2005;4:5.
- Yilmazer A. Cancer cell lines involving cancer stem cell populations respond to oxidative stress. Biotechnol Rep. 2017;17:24-30.
- Sander CS, Hamm F, Elsner P, Thiele JJ. Oxidative stress in malignant melanoma and non-melanoma skin cancer. Br J Dermatol. 2003;148:913-22.
- Villalta PC, Rocic P, Townsley MI. Role of MMP2 and MMP9 in TRPV4-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 2014;307:L652-9.
- Hong Z, Tian Y, Yuan Y, Qi M, Li Y, Du Y et al. Enhanced oxidative stress is responsible for TRPV4-induced neurotoxicity. Front Cell Neurosci. 2016;232.
- Özşimşek A, Nazıroğlu M. The involvement of TRPV4 on the hypoxia-induced oxidative neurotoxicity and apoptosis in a neuronal cell line: Protective role of melatonin. Neurotoxicology. 2021;87:136-48.
- Wu Q, Lu K, Zhao Z, Wang B, Liu H, Zhang S et al. Blockade of transient receptor potential vanilloid 4 enhances antioxidation after myocardial ischemia/reperfusion. Oxid Med Cell Longev. 2019;7283683.
- Nazıroğlu M. A novel antagonist of TRPM2 and TRPV4 channels: Carvacrol. Metab Brain Dis. 2022;37:711–28
- Liu X, Zhang P, Xie C, Sham KWY, Ng SSM, Chen Y et al. Activation of PTEN by inhibition of TRPV4 suppresses colon cancer development. Cell Death Dis. 2019;10:460
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Kanser Genetiği |
| Bölüm | Araştırma |
| Yazarlar | |
| Erken Görünüm Tarihi | 25 Eylül 2023 |
| Yayımlanma Tarihi | 30 Eylül 2023 |
| Kabul Tarihi | 11 Eylül 2023 |
| Yayımlandığı Sayı | Yıl 2023 Cilt: 48 Sayı: 3 |
