Myricetinin LNCaP Androjen Bağımlı Prostat Kanseri Hücreleri Üzerine Etkisinin Araştırılması

Prostat kanseri dünyada erkekler arasında en sık görülen ikinci kanser türüdür. Prostat kanserinin morbidite ve mortalitesi son zamanlarda artmıştır. Tedavisi için birçok alternatif yaklaşımlar geliştirilmeye çalışılsa da, prostat kanseri hala kötü prognoz sergilemekte ve yüksek ölüm oranları ile karşılaşılmaktadır. Myricetin, antikanser özelliği ile ilgi çeken doğal bir flavonoid bileşiktir. Yapılan in vitro ve in vivo çalışmalar myricetinin çeşitli mekanizmalar yoluyla prostat kanserini etkili bir şekilde inhibe ettiğini göstermektedir. Bu çalışmanın amacı artan dozlarda myricetin uygulamasının androjen reseptör bağımlı insan prostat kanser hücre hattı olan LNCaP hücrelerinin canlılığı üzerindeki etkilerini belirlemek ve apoptozla ilişkili BAX ve BCL2 genlerinin ekspresyon seviyelerini tespit etmektir. LNCaP hücreleri myricetinin 10 μM, 25 μM, 50 μM, 100 μM, 150 μM’lık konsantrasyonları ile 24 ve 48 saat süresince inkübe edilmiş ve hücre canlılığındaki değişimler 2,3-bis-(2-metoksi-4-nitro-5-sulfofenil)-2Htetrazolyum-5-karboksanilid (XTT) yöntemiyle belirlenerek IC50 değerleri hesaplanmıştır. BAX ve BCL2 gen ifadelerindeki değişimler ise Real-Time PCR metoduyla belirlenmiş ve elde edilen verilerin analizinde ΔΔCT metodu kullanılmıştır. Myricetinin uygulanan bütün dozlarda kontrole göre LNCaP hücre canlılığını azalttığı gösterilmiş olup IC50 değeri 24. saat için 123.76 μM, 48. saat için ise 84.79 μM olarak tespit edilmiştir. Ayrıca, myricetin uygulamasının apoptoz ilişkili BAX gen ifadesini istatistiksel olarak anlamlı bir düzeyde artırırken BCL2 gen ifadesini ise azalttığı görülmüştür. Myricetinin LNCaP hücrelerindeki antiproliferatif ve apoptotik etkileri daha detaylı olarak araştırılmalıdır.

Anahtar Kelimeler:

Myricetin, LNCaP hücre hattı, Apoptoz, BAX ve BCL2 Gen Ekspresyonu

Investigation of the Effect of Myricetin On Human Androgen Dependent Prostate Cancer Cells

Prostate cancer is the second most common type of cancer amongst men worldwide. The morbidity and mortality of prostate cancer has increased recently. Although many alternative approaches have been tried to be developed for the treatment, prostate cancer still has a poor prognosis and high mortality rates. Myricetin is a natural flavonoid compound of interest for its anticancer properties. In vitro and in vivo studies show that myricetin effectively impedes prostate cancer through several mechanisms. The aim of this study was to determine the effects of increasing doses of myricetin on the viability of LNCaP cells, an androgen receptor-dependent human prostate cancer cell line, and to determine the expression levels of the apoptosis-related BAX and BCL2 genes. LNCaP cells were incubated with myricetin at concentrations of 10 μM, 25 μM, 50 μM, 100 μM, 150 μM for 24 and 48 hours and alterations in cell viability were determined by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl) )-2Htetrazolium-5-carboxanilide (XTT) method and IC50 values were calculated. Changes in BAX and BCL2 gene expressions were investigated by Real-Time PCR and the data was analysed using ΔΔCT method. It was shown that myricetin decreased LNCaP cell viability at all doses used compared to the control, and the IC50 value was determined as 123.76 μM for the 24th hour and 84.79 μM for the 48th hour. In addition, it was observed that myricetin administration increased apoptosis associated BAX gene expression at a statistically significant level, while it was decreasing BCL2 gene expression. The antiproliferative and apoptotic effects of myricetin on LNCaP cells need to be investigated in more detail.

Keywords:

Myricetin, LNCaP cell line, Apoptosis, BAX and BCL2 Gene Expressions,

PDF

___

1. Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2022. CA Cancer J Clin. 2022;72(1):7-33.
2. Nelson WG, De Marzo AM, Isaacs WB. Prostate cancer. N Engl J Med. 2003;349(4):366-81.
3. Rebello RJ, Oing C, Knudsen KE, Loeb S, Johnson DC, Reiter RE, et al. Prostate cancer. Nature Reviews Disease Primers. 2021;7(1):9.
4. Armstrong CM, Gao AC. Dysregulated androgen synthesis and anti-androgen resistance in advanced prostate cancer. Am J Clin Exp Urol. 2021;9(4):292-300.
5. Sumanasuriya S, De Bono J. Treatment of Advanced Prostate Cancer-A Review of Current Therapies and Future Promise. Cold Spring Harb Perspect Med. 2018;8(6):a030635.
6. Matilla MA. Chapter 10 - Metabolic Responses of Plants Upon Different Plant–Pathogen Interactions. In: Ahmad P, Ahanger MA, Singh VP, Tripathi DK, Alam P, Alyemeni MN, editors. Plant Metabolites and Regulation Under Environmental Stress: Academic Press; 2018. p. 195-214.
7. Ganry O. Phytoestrogens and prostate cancer risk. Preventive medicine. 2005;41(1):1-6.
8. Haddad AQ, Venkateswaran V, Viswanathan L, Teahan SJ, Fleshner NE, Klotz LH. Novel antiproliferative flavonoids induce cell cycle arrest in human prostate cancer cell lines. Prostate Cancer and Prostatic Diseases. 2006;9(1):68-76.
9. Imran M, Saeed F, Hussain G, Imran A, Mehmood Z, Gondal TA, et al. Myricetin: A comprehensive review on its biological potentials. Food Sci Nutr. 2021;9(10):5854-68.
10. Zhu ML, Zhang PM, Jiang M, Yu SW, Wang L. Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells. BMC Complement Med Ther. 2020;20(1):209.
11. Berköz M, Yalın S, Özkan-Yılmaz F, Özlüer-Hunt A, Krośniak M, Francik R, et al. Protective effect of myricetin, apigenin, and hesperidin pretreatments on cyclophosphamide-induced immunosuppression. Immunopharmacology and Immunotoxicology. 2021;43(3):353-69.
12. Li Q, Tan Q, Ma Y, Gu Z, Chen S. Myricetin Suppresses Ovarian Cancer In Vitro by Activating the p38/Sapla Signaling Pathway and Suppressing Intracellular Oxidative Stress. Frontiers in Oncology. 2022;12:903394.
13. Tavsan Z, Kayali HA. Flavonoids showed anticancer effects on the ovarian cancer cells: Involvement of reactive oxygen species, apoptosis, cell cycle and invasion. Biomedicine & Pharmacotherapy. 2019;116:109004.
14. Xu Y, Xie Q, Wu S, Yi D, Yu Y, Liu S, et al. Myricetin induces apoptosis via endoplasmic reticulum stress and DNA double-strand breaks in human ovarian cancer cells. Mol Med Rep. 2016;13(3):2094-100.
15. Zhang S, Wang L, Liu H, Zhao G, Ming L. Enhancement of recombinant myricetin on the radiosensitivity of lung cancer A549 and H1299 cells. Diagn Pathol. 2014;9:68.
16. Wang L, Feng J, Chen X, Guo W, Du Y, Wang Y, et al. Myricetin enhance chemosensitivity of 5-fluorouracil on esophageal carcinoma in vitro and in vivo. Cancer Cell Int. 2014;14:71.
17. Ha TK, Jung I, Kim ME, Bae SK, Lee JS. Anti-cancer activity of myricetin against human papillary thyroid cancer cells involves mitochondrial dysfunction–mediated apoptosis. Biomedicine & Pharmacotherapy. 2017;91:378-84.
18. Jiang M, Zhu M, Wang L, Yu S. Anti-tumor effects and associated molecular mechanisms of myricetin. Biomedicine & Pharmacotherapy. 2019;120:109506.
19. Jose J, Dhanya AT, Haridas KR, Sumesh Kumar TM, Jayaraman S, Variyar EJ, et al. Structural characterization of a novel derivative of myricetin from Mimosa pudica as an anti-proliferative agent for the treatment of cancer. Biomedicine & Pharmacotherapy. 2016;84:1067-77.
20. Sajedi N, Homayoun M, Mohammadi F, Soleimani M. Myricetin Exerts its Apoptotic Effects on MCF-7 Breast Cancer Cells through Evoking the BRCA1-GADD45 Pathway. Asian Pac J Cancer Prev. 2020;21(12):3461-8.
21. Ji A, Hu L, Ma D, Qiang G, Yan D, Zhang G, et al. Myricetin Induces Apoptosis and Protective Autophagy through Endoplasmic Reticulum Stress in Hepatocellular Carcinoma. Evid Based Complement Alternat Med. 2022;2022:3115312.
22. Li HG, Chen JX, Xiong JH, Zhu JW. Myricetin exhibits anti-glioma potential by inducing mitochondrial-mediated apoptosis, cell cycle arrest, inhibition of cell migration and ROS generation. J BUON. 2016;21(1):182-90.
23. Li Y, Cui SX, Sun SY, Shi WN, Song ZY, Wang SQ, et al. Chemoprevention of intestinal tumorigenesis by the natural dietary flavonoid myricetin in APCMin/+ mice. Oncotarget. 2016;7(37):60446-60.
24. Agraharam G, Girigoswami A, Girigoswami K. Myricetin: a Multifunctional Flavonol in Biomedicine. Current Pharmacology Reports. 2022;8.
25. Ni F, Gong Y, Li L, Abdolmaleky HM, Zhou JR. Flavonoid ampelopsin inhibits the growth and metastasis of prostate cancer in vitro and in mice. PloS one. 2012;7(6):e38802.
26. Ye C, Zhang C, Huang H, Yang B, Xiao G, Kong D, et al. The Natural Compound Myricetin Effectively Represses the Malignant Progression of Prostate Cancer by Inhibiting PIM1 and Disrupting the PIM1/CXCR4 Interaction. Cellular Physiology and Biochemistry. 2018;48(3):1230-44.
27. Zheng AW, Chen YQ, Zhao LQ, Feng JG. Myricetin induces apoptosis and enhances chemosensitivity in ovarian cancer cells. Oncol Lett. 2017;13(6):4974-8.
28. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71(3):209-49.
29. Zorlu F, Zorlu R, Divrik RT, Eser S, Yorukoglu K. Prostate cancer incidence in Turkey: an epidemiological study. Asian Pac J Cancer Prev. 2014;15(21):9125-30.
30. Klein EA, Thompson IM. Chemoprevention of prostate cancer: an updated view. World journal of urology. 2012;30(2):189-94.
31. Huang J, Plass C, Gerhauser C. Cancer chemoprevention by targeting the epigenome. Curr Drug Targets. 2011;12(13):1925-56.
32. Xu R, Zhang Y, Ye X, Xue S, Shi J, Pan J, et al. Inhibition effects and induction of apoptosis of flavonoids on the prostate cancer cell line PC-3 in vitro. Food Chemistry. 2013;138(1):48-53.
33. Liu J-S, Fang W-K, Yang S-M, Wu M-C, Chen T-J, Chen C-M, et al. Natural product myricetin is a pan-KDM4 inhibitor which with poly lactic-co-glycolic acid formulation effectively targets castration-resistant prostate cancer. Journal of Biomedical Science. 2022;29(1):29.
34. Jin C, Wu S, Lu X, Liu Q, Zhang L, Yang J, et al. Conditioned medium from actinomycin D-treated apoptotic cells induces mitochondria-dependent apoptosis in bystander cells. Toxicology letters. 2012;211(1):45-53.
35. Zhang H, Heim J, Meyhack B. Redistribution of Bax from Cytosol to Membranes Is Induced by Apoptotic Stimuli and Is an Early Step in the Apoptotic Pathway. Biochemical and Biophysical Research Communications. 1998;251(2):454-9.
36. Ola MS, Nawaz M, Ahsan H. Role of Bcl-2 family proteins and caspases in the regulation of apoptosis. Molecular and cellular biochemistry. 2011;351(1-2):41-58.
37. Martinou JC, Youle RJ. Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Developmental cell. 2011;21(1):92-101.
38. Donovan M, Cotter TG. Control of mitochondrial integrity by Bcl-2 family members and caspase-independent cell death. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 2004;1644(2):133-47.
39. Zhang XH, Chen SY, Tang L, Shen YZ, Luo L, Xu CW, et al. Myricetin induces apoptosis in HepG2 cells through Akt/p70S6K/bad signaling and mitochondrial apoptotic pathway. Anti-cancer agents in medicinal chemistry. 2013;13(10):1575-81.
40. Jiao D, Zhang XD. Myricetin suppresses p21-activated kinase 1 in human breast cancer MCF-7 cells through downstream signaling of the β-catenin pathway. Oncology reports. 2016;36(1):342-8.
41. Kim ME, Ha TK, Yoon JH, Lee JS. Myricetin Induces Cell Death of Human Colon Cancer Cells via BAX/BCL2-Dependent Pathway. Anticancer Research. 2014;34(2):701.
42. Sun W, Tao Y, Yu D, Zhao T, Wu L, Yu W, et al. Myricetin exerts potent anticancer effects on human skin tumor cells. Tropical Journal of Pharmaceutical Research. 2018;17:1067-72.