Determination of Antiproliferative and Necroptotic Effects of Chalcone Compounds on Prostate Cancer Cells

Year 2023, , 1552 - 1561, 01.09.2023

Elif Erturk , Ayşen Sağnak , Yaren Yıldız , Oğuzhan Akgün , Demet Coşkun , Ferda Arı

https://doi.org/10.21597/jist.1298265

Abstract

Cancer is a multistage fatal disease that occurs with genetic and environmental conditions and uncontrolled division and proliferation of cells. Prostate cancer is one of the malignancies that affects men and contributes significantly to the increasing mortality rates in men globally. Due to a long disease history, genetic-phenotypic diversity, and uncertainty in the clinical progression of patients in prostate cancer, the necessity of developing new approaches arises. Chalcones are pharmacologically active compounds found in plants. It has been observed that natural or synthetic chalcone derivatives have anti-cancer activity in cancer cells. In the current study, the anti-cancer and anti-proliferative effects of two synthesized and characterized chalcone derivatives (Compound 1 and Compound 2) were investigated in human prostate cancer cell lines (LNCaP and PC-3). The anti-proliferative effect of the compounds on cell viability was assessed by SRB viability assay after 48 hours of treatment. Fluorescent staining (Hoechst 33342+Annexin-V+Propidium iodide) method was used to determine the cell death mode responsible for the cytotoxic effects of chalcone compounds, and RT-PCR analysis was performed to determine the changes in gene expressions. Anti-proliferative effects of the compounds were detected in LNCaP and PC-3 cells in a dose- and time-dependent manner. It was determined by triple fluorescent staining that the compounds induced secondary apoptosis in LNCaP and PC-3 cells. Significant increases in expression levels of cell death pathway genes BCL-2, MLKL, FAS and PARP were determined. In the light of the results obtained, it was concluded that Compound 1 and Compound 2 showed anti-proliferative effect and induced necroptosis in prostate cancer.

Keywords

Prostate cancer, Chalcones, Antiproliferative, Anticancer, Necroptosis

Project Number

FLO-2022-821

Kalkon Bileşiklerinin Prostat Kanseri Hücrelerinde Antiproliferatif ve Nekroptotik Etkilerinin Belirlenmesi

Abstract

Kanser, genetik ve çevresel koşullar ile hücrelerin kontrolsüz bölünmesi ve çoğalmasıyla ortaya çıkan çok basamaklı ölümcül bir hastalıktır. Prostat kanseri, erkekleri etkileyen ve küresel olarak erkeklerde artan ölüm oranlarına önemli ölçüde katkıda bulunan malignitelerden biridir. Prostat kanserinde, uzun bir hastalık geçmişi, genetik-fenotipik çeşitlilik ve hastaların klinik ilerlemesindeki belirsizlik nedeniyle yeni yaklaşımların geliştirilmesi gerekliliği ortaya çıkmaktadır. Kalkonlar, bitkilerde bulunan, farmakolojik aktif bileşiklerdir. Doğal ya da sentetik kalkon türevlerinin kanser hücrelerinde anti-kanser aktiviteye sahip olduğu gözlenmiştir. Mevcut çalışmada, sentezi ve karakterizasyonu yapılmış iki kalkon türevinin (Bileşik 1 ve Bileşik 2) anti-kanser ve anti-proliferatif etkileri insan prostat kanseri hücre soylarında (LNCaP ve PC-3) araştırıldı. Bileşiklerin hücre canlılığı üzerindeki anti-proliferatif etkisi 48 saatlik tedavi sonrası SRB canlılık analizi ile değerlendirildi. Kalkon bileşiklerinin sitotoksik etkilerinden sorumlu hücre ölüm modunu belirlemek amacıyla floresan boyama (Hoechst 33342+Anneksin-V+Propidyum iyodür) yöntemi ve gen ekspresyonlarında meydana getirdiği değişiklikleri saptamak için de RT-PCR analizi gerçekleştirildi. Bileşiklerin, LNCaP ve PC-3 hücrelerinde doza ve zamana bağlı olarak anti-proliferatif etkileri saptandı. Üçlü floresan boyama sonucu ile bileşiklerin LNCaP ve PC-3 hücrelerinde sekonder apoptozisi teşvik ettiği tespit edildi. Hücre ölümü yolak genleri BCL-2, MLKL, FAS ve PARP ekspresyon seviyelerinde anlamlı artışlar belirlendi. Elde edilen sonuçların ışığında, Bileşik 1 ve Bileşik 2’nin prostat kanserinde anti-proliferatif etki gösterdiği ve nekroptozisi indüklediği sonucuna varıldı.

Keywords

Prostat kanseri, Kalkonlar, Antiproliferatif, Antikanser, Nekroptozis

Supporting Institution

Bursa Uludağ Üniversitesi

Project Number

FLO-2022-821

References

• Abou-Zied, H. A., Youssif, B. G., Mohamed, M. F., Hayallah, A. M., & Abdel-Aziz, M. (2019). EGFR inhibitors and apoptotic inducers: Design, synthesis, anticancer activity and docking studies of novel xanthine derivatives carrying chalcone moiety as hybrid molecules. Bioorganic chemistry, 89, 102997.
• Alioglu, I., Cinar-Asa, S., Ari, F., Coskun, D. (2023). Benzofuran substituted chalcone derivatives trigger apoptotic cell death through extrinsic pathway in human lung and breast cancer cells. Iranian Journal of Science and Technology Transactions A: Science,
• Arif, R., Rana, M., Yasmeen, S., Khan, M. S., Abid, M., & Khan, M. S. (2020). Facile synthesis of chalcone derivatives as antibacterial agents: Synthesis, DNA binding, molecular docking, DFT and antioxidant studies. Journal of Molecular Structure, 1208, 127905.
• Bach, C., Pisipati, S., Daneshwar, D., Wright, M., Rowe, E., Gillatt, D., & Koupparis, A. (2014). The status of surgery in the management of high-risk prostate cancer. Nature Reviews Urology, 11(6), 342-351.
• Beytur, A., Tekin, Ç., Çalışkan, E., Tekin, S., Koran, K., Görgülü, A. O., & Sandal, S. (2022). Hexa-substituted cyclotriphosphazene derivatives containing hetero-ring chalcones: Synthesis, in vitro cytotoxic activity and their DNA damage determination. Bioorganic Chemistry, 127, 105997.
• Chen, J., Zhang, D., Yan, W., Yang, D., & Shen, B. (2013). Translational bioinformatics for diagnostic and prognostic prediction of prostate cancer in the next-generation sequencing era. BioMed research international, 2013.
• Coşkun, D., & Ahmedzade, M. (2014). Synthesis of some acylated 2-pyrazoline and chalcone derivatives. Research on Chemical Intermediates, 40, 1193-1199.
• Coşkun, D., Tekin, S., Sandal, S., & Coşkun, M. F. (2016). Synthesis, characterization, and anticancer activity of new benzofuran substituted chalcones. Journal of Chemistry, 2016.
• Coskun, D., Erkisa, M., Ulukaya, E., Coskun, M. F., & Ari, F. (2017). Novel 1-(7-ethoxy-1-benzofuran-2-yl) substituted chalcone derivatives: synthesis, characterization and anticancer activity. European journal of medicinal chemistry, 136, 212-222.
• Dong, N., Liu, X., Zhao, T., Wang, L., Li, H., Zhang, S., & Yang, B. (2018). Apoptosis-inducing effects and growth inhibitory of a novel chalcone, in human hepatic cancer cells and lung cancer cells. Biomedicine & Pharmacotherapy, 105, 195-203.
• Ducki, S. (2007). The development of chalcones as promising anticancer agents. IDrugs, 10(1), 42.
• Erturk, E., Tuna, G., Coskun, D., & Ari, F. (2023). Investigation of Anti-Cancer Activity of Newly Synthesized 2, 4-pentadien-1-one Derivative Containing Benzofuran in Human Lung and Colon Cancer Cells.
• Escobar, S. J. D. M., Fong, G. M., Winnischofer, S. M., Simone, M., Munoz, L., Dennis, J. M., & Witting, P. K. (2019). Anti-proliferative and cytotoxic activities of the flavonoid isoliquiritigenin in the human neuroblastoma cell line SH-SY5Y. Chemico-Biological Interactions, 299, 77-87.
• Hanahan, D., & Weinberg, R. A. (2011). Hallmarks of cancer: the next generation. cell, 144(5), 646-674.
• Hanahan, D. (2022). Hallmarks of cancer: new dimensions. Cancer discovery, 12(1), 31-46.
• He, W., Wang, Q., Srinivasan, B., Xu, J., Padilla, M. T., Li, Z., & Lin, Y. (2014). A JNK-mediated autophagy pathway that triggers c-IAP degradation and necroptosis for anticancer chemotherapy. Oncogene, 33(23), 3004-3013.
• Hussaini, S. M. A., Yedla, P., Babu, K. S., Shaik, T. B., Chityal, G. K., & Kamal, A. (2016). Synthesis and biological evaluation of 1, 2, 3‐triazole tethered pyrazoline and chalcone derivatives. Chemical biology & drug design, 88(1), 97-109.
• Hussain, S., Singh, A., Nazir, S. U., Tulsyan, S., Khan, A., Kumar, R., ... & Mehrotra, R. (2019). Cancer drug resistance: a fleet to conquer. Journal of Cellular Biochemistry, 120(9), 14213-14225.
• Kim, S. J., & Li, J. (2013). Caspase blockade induces RIP3-mediated programmed necrosis in Toll-like receptor-activated microglia. Cell death & disease, 4(7), e716-e716.
• Özen, F., Günel, A., & Baran, A. (2018). DNA-binding, enzyme inhibition, and photochemical properties of chalcone-containing metallophthalocyanine compounds. Bioorganic chemistry, 81, 71-78.
• Panche, A. N., Diwan, A. D., & Chandra, S. R. (2016). Flavonoids: an overview. Journal of nutritional science, 5, e47.
• Sezgin, C. (2010). Kanserde bitkilerle tedavide örnek uygulamalar. Bitkilerle Tedavi, 73.
• Sharma, V., Chaudhary, A., Arora, S., Saxena, A. K., & Ishar, M. P. S. (2013). β-Ionone derived chalcones as potent antiproliferative agents. European Journal of Medicinal Chemistry, 69, 310-315.
• Siegel, R. L., Miller, K. D., Wagle, N. S., & Jemal, A. (2023). Cancer statistics, 2023. CA: a cancer journal for clinicians, 73(1), 17-48.
• Sung, H., Ferlay, J., Siegel, R. L., Laversanne, M., Soerjomataram, I., Jemal, A., & Bray, F. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians, 71(3), 209-249.
• Termini, D., Den Hartogh, D. J., Jaglanian, A., & Tsiani, E. (2020). Curcumin against prostate cancer: current evidence. Biomolecules, 10(11), 1536.
• Ulukaya, E., Acilan, C., & Yilmaz, Y. (2011). Apoptosis: why and how does it occur in biology?. Cell biochemistry and function, 29(6), 468-480.
• Zhuang, C., Zhang, W., Sheng, C., Zhang, W., Xing, C., & Miao, Z. (2017). Chalcone: a privileged structure in medicinal chemistry. Chemical reviews, 117(12), 7762-7810.
• Wan, M., Xu, L., Hua, L., Li, A., Li, S., Lu, W., ... & Jiao, P. (2014). Synthesis and evaluation of novel isoxazolyl chalcones as potential anticancer agents. Bioorganic chemistry, 54, 38-43.