ORSİNOL’ÜN SW480 İNSAN KOLOREKTAL KANSER HÜCRELERİNDE PROLİFERASYON VE APOPTOZ ÜZERİNE ETKİLERİ

Year 2023, , 375 - 382, 20.05.2023

Betül Yanık Filiz Bakar Ateş

https://doi.org/10.33483/jfpau.1228071

Abstract

Amaç: Kolorektal kanser, her yıl dünya çapında hızla artan bir hastalıktır. Teşhis konulmuş hastaların neredeyse yarısı her yıl bu hastalıktan hayatını kaybetmektedir. Kolorektal kanserin tedavisi için uygulanan geleneksel tedavi yöntemleri olan cerrahi, radyoterapi ve mevcut kemoterapi seçenekleri de dahil olmak üzere tedavide kullanılan yöntemlerin etkinliği düşüktür ve çok fazla yan etkileri bulunmaktadır. Tüm bu problemler nedeni ile kolorektal kanserin tedavisi için yeni ajanlar geliştirmenin önemi giderek artmaktadır. Orsinol, likenlerden izole edilen bir sekonder metabolit olup, söz konusu bileşiğin antioksidan, antimikrobiyal ve antidepresan aktivitesine yönelik bulgular mevcuttur. Son yıllarda bileşiğin antikanser etkinliğine dair araştırmalar da literatürde yer almaya başlamıştır. Bu çalışmada, orsinolün insan SW480 kolorektal kanser hücrelerinde hücre proliferasyonu ve apoptoz üzerine etkinliğinin araştırılması amaçlanmıştır.
Gereç ve Yöntem: Çalışma kapsamında SW480 insan kolorektal kanser hücreleri kullanılmış ve DMEM besiyerinde kültüre edilmiştir. Orsinol, dimetilsülfoksit ile çözülerek stok çözeltisi hazırlanmış ve hücrelere 1-25 mM konsantrasyon aralığında uygulanmıştır. Orsinolün hücre canlılığı üzerine etkisi MTT testi ile belirlenmiştir. Bileşiğin apoptotik etkinliği Annexin V bağlanma analizi ile Muse Hücre Analiz cihazı kullanılarak değerlendirilmiştir.
Sonuç ve Tartışma: MTT analiz sonuçları, orsinolün 5 mM ve üzerinde hücre canlılığını doza bağlı olarak anlamlı şekilde azalttığını gösterdi (p<0.05). Kontrol grubunda hücre canlılığı 100.00±6.14% iken, 25 mM orsinol uygulanan hücrelerde canlılık 12.50±0.65% olarak belirlendi (p<0.0001). Annexin V bağlanma analizi bulgularına göre erken apoptotik hücre popülasyonu 25 mM orsinol uygulanan grupta 12.06±1.22% iken, kontrol grubunda 0.60±0.11% olarak belirlendi. Çalışmadan elde edilen bulgular, orsinol’ün, SW480 kolorektal kanser hücreleri üzerinde yüksek konsantrasyonda sitotoksik etkili olduğunu göstermiş olup, daha ileri çalışmalar ile bileşiğin etkinliğinin artırılmasına ve etki mekanizmasının aydınlatılmasına ihtiyaç duyulmaktadır.

Keywords

Apoptoz, kolorektal kanser, orsinol, proliferasyon, SW480

THE EFFECTS OF ORCINOL ON PROLIFERATION AND APOPTOSIS OF SW480 HUMAN COLORECTAL CANCER CELLS

Abstract

Objective: Colorectal cancer is a rapidly increasing disease worldwide, and almost half of the diagnosed patients die from this disease each year. The methods used in the treatment of colorectal cancer, including traditional treatment methods such as surgery, radiotherapy and current chemotherapy options, have low effectiveness and have many side effects. Because of all these problems, the importance of developing new agents for the treatment of colorectal cancer is increasing. Orsinol is a secondary metabolite isolated from lichens, and there are findings regarding the antioxidant, antimicrobial and antidepressant activity of the compound. In recent years, research on the anticancer activity of the compound has also started to take place in the literature. In this study, it was aimed to investigate the efficacy of orcinol on cell proliferation and apoptosis in human SW480 colorectal cancer cells.
Material and Method: Within the scope of the study, SW480 human colorectal cancer cells were used and cultured in DMEM medium. Orcinol was dissolved with dimethylsulfoxide to prepare a stock solution and applied to the cells in a concentration range of 1-25 mM. The effect of orcinol on cell viability was determined by MTT test. The apoptotic activity of the compound was evaluated with Annexin V binding assay using the Muse Cell Analyzer.
Result and Discussion: The results of MTT analysis showed that orcinol significantly decreased cell viability at 5 mM and above (p<0.05). While cell viability was 100.00±6.14% in the control group, it was determined as 12.50±0.65% in cells treated with 25 mM orcinol (p<0.0001). According to Annexin V binding analysis findings, the early apoptotic cell population was 12.06±1.22% in the 25 mM orcinol treated group, while it was 0.60±0.11% in the control group. The findings obtained from the study showed that that orcinol has a cytotoxic effect at high concentration on SW480 colorectal cancer cells, and further studies are needed to increase the efficiency of the compound and to elucidate its mechanism of action.

Keywords

Apoptosis, colorectal cancer, orcinol, proliferation, SW480.

References

• 1. 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. [CrossRef]
• 2. Bray, F., Ferlay, J., Soerjomataram, I., Siegel, R.L., Torre, L.A., Jemal, A. (2018). Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians, 68(6), 394-424. [CrossRef]
• 3. Arnold, M., Sierra, M.S., Laversanne, M., Soerjomataram, I., Jemal, A., Bray, F. (2017). Global patterns and trends in colorectal cancer incidence and mortality. Gut, 66(4), 683-691. [CrossRef]
• 4. Turkey Cancer Statistics Web site. (2017), from https://hsgm.saglik.gov.tr/tr/kanser-istatistikleri/yillar/2017-turkiye-kanser-i-statistikleri.html Accessed date: 29.11.2022.
• 5. Zhang, J., Jiang, Y., Zhu, J., Wu, T., Ma, J., Du, C., Chen, S., Li, T., Han, J., Wang, X. (2017). Overexpression of long non-coding RNA colon cancer-associated transcript 2 is associated with advanced tumor progression and poor prognosis in patients with colorectal cancer. Oncology Letters, 14(6), 6907-6914. [CrossRef]
• 6. Dekker, E., Tanis, P.J., Vleugels, J.L.A., Kasi, P.M., Wallace, M.B. (2019). Colorectal cancer. Lancet, 394(10207), 1467-1480. [CrossRef]
• 7. George, J., Rockall, T. (2020). Surgery for colorectal cancer. Surgery, 38(1), 32-37. [CrossRef]
• 8. Wang, Y., Cheng, X., Wang, P., Wang, L., Fan, J., Wang, X., Liu, Q. (2014). Investigating migration inhibition and apoptotic effects of fomitopsis pinicola chloroform extract on human colorectal cancer SW-480 cells. Plos One, 9(7), e101303. [CrossRef]
• 9. Sharifi-Rad, J., Ozleyen, A., Boyunegmez Tumer, T., Oluwaseun Adetunji, C., El Omari, N., Balahbib, A., Taheri, Y., Bouyahya, A., Martorell, M., Martins, N., Cho, W.C. (2019). Natural Products and Synthetic Analogs as a Source of Antitumor Drugs. Biomolecules, 9(11), 679. [CrossRef]
• 10. Newman, D.J., Crag, G.M. (2016). Natural products as sources of new drugs from 1981 to 2014. Journal of Natural Products,79(3), 629-661. [CrossRef]
• 11. Guo, J., Li, Z.L., Wang, A.L., Liu, X.Q., Wang, J., Guo, X., Jing, Y.K, Hua H.M. (2011). Three new phenolic compounds from the lichen Thamnolia vermicularis and their antiproliferative effects in prostate cancer cells. Planta Medica, 77(18), 2042-2046. [CrossRef]
• 12. Hladyszowski, J., Zubik, L., Kozubek, A. (1998). Quantum mechanical and experimental oxidation studies of pentadecylresorcinol, olivetol, orcinol and resorcinol. Free Radical Research, 28(4), 359-368. [CrossRef]
• 13. Ivanova, V., Backor, M., Dahse, H.M., Graefe, U. (2010). Molecular structural studies of lichen substances with antimicrobial, antiproliferative, and cytotoxic effects from Parmelia subrudecta. Preparative Biochemistry and Biotechnology, 40(4), 377-388. [CrossRef]
• 14. Lopes T.I.B., Coelho, R.G., Yoshida, N.C., Honda, N.K., (2008). Radical-scavenging activity of orsellinates. Chemical and Pharmaceutical Bulletin, 56(11), 1551-1554. [CrossRef]
• 15. Ge, J.F., Gao, W.C., Cheng, W.M., Lu, W.L., Tang, J., Peng, L., Li, N., Chen, F.H., (2014). Orcinol glucoside produces antidepressant effects by blocking the behavioural and neuronal deficits caused by chronic stress. European Neuropsychopharmacology, 24(1), 172-180. [CrossRef]
• 16. Murakami, Y., Kawata, A., Ito, S., Katayama, T., Fujisawa, S. (2015) The radical scavenging activity and cytotoxicity of resveratrol, orcinol and 4-Allylphenol and their inhibitory effects on Cox-2 gene expression and nf-ĸb activation in RAW264.7 cells stimulated with porphyromonas gingivalis-fimbriae. In Vivo, 29(3), 341-350.
• 17. Nahak, P., Gajbhiye, R.L., Karmakar, G., Guha, P., Roy, B., Besra, S.E., Bikov, A.G., Akentiev, A.V., Noskov, B.A., Nag, K., Jaisankar, P., Panda A.K. (2018). Orcinol glucoside loaded polymer-lipid hybrid nanostructured lipid carriers: potential cytotoxic agents against gastric, colon and hepatoma carcinoma cell lines. Pharmaceutical Research, 35(10), 198. [CrossRef]
• 18. Jemal, A., Siegel, R., Ward, E., Hao, Y., Xu, J., Murray, T., Thun, M.J. (2008). Cancer statistics, 2008. CA: A Cancer Journal for Clinicians, 58(2), 71-96. [CrossRef]
• 19. Oehler, C., Ciernik, I.F. (2006). Radiation therapy and combined modality treatment of gastrointestinal carcinomas Cancer Treatment Reviews, 32(2), 119-138. [CrossRef]
• 20. Matsui, T., Omura, K., Kawakami, K., Morita, S., Sakamoto, J. (2006). Genotype of thymidylate synthase likely to affect efficacy of adjuvant 5-FU based chemotherapy in colon cancer. Oncology Reports, 16(5), 1111-1115. [CrossRef]
• 21. Bakar, F. (2016). Cucurbitacin B Enhances the Anticancer Effect of Imatinib Mesylate Through Inhibition of MMP-2 Expression in MCF-7 and SW480 Tumor Cell Lines. Anti-Cancer Agents in Medicinal Chemistry, 16(6), 747-754. [CrossRef]
• 22. Bakar-Ates, F., Ozkan, E. (2019). The combined treatment of brassinin and imatinib synergistically downregulated the expression of MMP-9 in SW480 colon cancer cells. Phytotherapy Research, 33(2), 397-402. [CrossRef]
• 23. Molnár, K., Farkas, E. (2010). Current results on biological activities of lichen secondary metabolites: a review. Zeitschrift für Naturforschung C, 65(3-4), 157-173. [CrossRef]
• 24. Manojlović, N.T., Vasiljević, P., Jusković, M., Najman, S., Janković, S., Milenković-Andjelković, A. (2010). HPLC analysis and cytotoxic potential of extracts from the lichen, Thamnolia vermicularis var. subuliformis. Journal of Medicinal Plants Research, 4(9), 817-823.
• 25. Shukla, V., Joshi, G.P., Rawat, M.S.M. (2010). Lichens as a potential natural source of bioactive compounds: a review. Phytochemistry Reviews, 9(2), 303-314. [CrossRef]
• 26. Manojlovic, N.T., Solujic, S., Sukdolak, S. (2002). Antimicrobial activity of an extract and anthraquinones from caloplaca schaereri. The Lichenologist, 34(1), 83-85. [CrossRef]
• 27. Hidalgo, M.E., Fernández, E., Quilhot, W., Lissi, E. (1994). Antioxidant activity of depsides and depsidones. Phytochemistry, 37(6), 1585-1587. [CrossRef]
• 28. Tang, S.Y., Whiteman, M., Peng, Z.F., Jenner, A., Yong, E.L., Halliwell, B. (2004). Characterization of antioxidant and antiglycation properties and isolation of active ingredients from traditional Chinese medicines. Free Radical Biology and Medicine, 36(12), 1575-1587. [CrossRef]
• 29. Wu, Q., Fu, D.X., Hou, A.J., Lei, G.Q., Liu, Z.J., Chen, J.K., Zhou, T.S. (2005). Antioxidative phenols and phenolic glycosides from Curculigo orchioides. Chemical and Pharmaceutical Bulletin, 53(8), 1065-1067. [CrossRef]
• 30. Bafna, A.R., Mishra, S.H. (2006). Immunostimulatory effect of methanol extract of Curculigo orchioides on immunosuppressed mice. Journal of Ethnopharmacology, 104(1-2), 1-4. [CrossRef]
• 31. Taborga, L., Espinoza, L., Moller, A., Carrasco, H., Cuellar, M., Villena, J. (2016). Antiproliferative effect and apoptotic activity of linear geranylphenol derivatives from phloroglucinol and orcinol. Chemico-Biological Interactions, 247, 22-29. [CrossRef]
• 32. Thadhani, V.M., Mesaik, M.A., Asif, M., Karunaratne, V., Choudhary, I.M. (2015). Immunomodulatory activities of some common lichen metabolites. International Journal of Pharmacy and Pharmaceutical Sciences, 7(11), 144-147.
• 33. Biskup, I., Zaczynska, E., Krauze-Baranowska, M., Fecka, I. (2017). Evaluation of cytotoxicity of 5-n-alkylresorcinol homologs and fraction on mouse fibroblast cell line L929. European Food Research and Technology, 243(7), 1137-1148. [CrossRef]