Evaluation of anticancerogenic effect of flavonoid rich Verbascum gypsicola Vural & Aydoğdu methanolic extract against SH-SY5Y cell line
Year 2024,
, 1 - 12, 30.06.2024
Seda Şirin
Abstract
Neuroblastoma (NB) is an embryonal neoplasm affecting the autonomic branch of the nervous system; it is the most commonly detected cancer type in children. NBs affecting children mostly present with metastatic disease that is hardly treatable with intensive multimodal therapy and portends a poor prognosis. Therefore, the likelihood of children with high-risk NB relapse remains extremely high, which calls for urgent action to discover novel treatment options to improve survival. Assessing the anti-cancer properties of known natural compounds may offer novel therapeutic options against NB. In this study we aimed to investigate the anti-cancer properties of the Verbacum gypsicola methanol extract (VGME) rich in flavonoids on SH-SY5Y cell line. For this purpose, we used LC-MS analysis to investigate the flavonoid composition of VGME, MTT analysis to investigate its effect on cell viability, and flow cytometry and qRT-PCR analyses to investigate its effect on apoptosis. VGME had a high flavonoid content. Its IC50 dose was 50 μg/mL at 48 hours. It significantly increased intracellular ROS level, apoptotic cells’ percentage, and mitochondrial disruption. The capacity of VGME to block cancer growth via an intrinsic apoptotic route implies that it might be a classic option for anticancer drug creation.
Ethical Statement
Not applicable.
Supporting Institution
This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.
Thanks
I would like to thank Prof. Dr. Belma Aslım for providing Verbascum gypsicola Vural & Aydoğdu methanolic extract.
References
- Abotaleb, M., Samuel, S. M., Varghese, E., Varghese, S., Kubatka, P., Liskova, A., Büsselberg, D. (2018). Flavonoids in cancer and apoptosis. Cancers, 11(1), 28.
https://doi.org/10.3390/cancers11010028.
- Afifi, M. S., Ahmed, M. M., Pezzuto, J. M., Kinghornt, A. D. (1993). Cytotoxic flavonolignans and flavones from Verbascum sinaiticum leaves. Phytochemistry, 34(3), 839-841.
https://doi.org/10.1016/0031-9422(93)85369-3.
- Alipieva, K. I., Orhan, I. E., Cankaya, I. I. T., Kostadinova, E. P., Georgiev, M. I. (2014). Treasure from garden: Chemical profiling, pharmacology and biotechnology of mulleins. Phytochemistry Reviews, 13, 417-444. https://doi.org/10.1007/s11101-014-9361-5.
- Alkowni, R., Jaradat, N., Fares, S. (2023). Total phenol, flavonoids, and tannin contents, antimicrobial, antioxidant, vital digestion enzymes inhibitory and cytotoxic activities of Verbascum fruticulosum. European Journal of Integrative Medicine, 60, 102256.https://doi.org/10.1016/j.eujim.2023.102256.
- Amini, S., Hassani, A., Alirezalu, A., Maleki, R. (2022). Phenolic and flavonoid compounds and antioxidant activity in flowers of nine endemic Verbascum species from Iran. Journal of the Science of Food and Agriculture, 102(8), 3250-3258.https://doi.org/10.1002/jsfa.11667.
- Amiri, M. M., Garnida, Y., Almulla, A. F., Abduljabbar, A. S., Jalil, A. T., Mazaheri, Y., Ebrahimi, Y., Shariatifar, N. (2023). Herbal therapy for hemorrhoids: An overview of medicinal plants affecting hemorrhoids. Advancements in Life Sciences, 10(1), 22-28. https://doi.org/10.1003/s101-017-2228-5.
- Anoushirvani, A. A., Jafarian Yazdi, A., Amirabadi, S., Asouri, S. A., Shafabakhsh, R., Sheida, A., Khabr, M. S. H., Jafari, A., Zadeh, S. S. T., Hamblin, M. R., Kalantari, L., Zavareh, S. A. T., Mirzaei, H. (2023). Role of non-coding RNAs in neuroblastoma. Cancer Gene Therapy, 1-19. https://doi.org/10.1038/s41417-023-00623-0.
- Ataur Rahman, M., Kim, N. H., Yang, H., Huh, S. O. (2012). Angelicin induces apoptosis through intrinsic caspase-dependent pathway in human SH-SY5Y neuroblastoma cells. Molecular and Cellular Biochemistry, 369, 95-104. https://doi.org/10.1007/s11010-012-1372-1.
- Aydin, Ç., Rakhimzhanova, A., Kilinçarslan, Ö., Mammadov, R. (2020). Antioxidant and phenolic characterization with HPLC of various extract of Verbascum glomeratum Linneus. Journal of The Chemical Society of Pakistan, 42(2), 222-227. https://doi.org/10.52568/000639/JCSP/42.02.2020.
- Cai, X., Ye, T., Liu, C., Lu, W., Lu, M., Zhang, J., Wang, M., Cao, P. (2011). Luteolin induced G2 phase cell cycle arrest and apoptosis on non-small cell lung cancer cells. Toxicology in Vitro, 25(7), 1385-1391.
https://doi.org/10.1016/j.tiv.2011.05.009.
- Cetraro, P., Plaza-Diaz, J., MacKenzie, A., Abadía-Molina, F. (2022). A review of the current impact of inhibitors of apoptosis proteins and their repression in cancer. Cancers, 14(7), 1671.
https://doi.org/10.3390/cancers14071671.
- Chen, J., Chen, J., Li, Z., Liu, C., Yin, L. (2014b). The apoptotic effect of apigenin on human gastric carcinoma cells through mitochondrial signal pathway. Tumor Biology, 35, 7719-7726.
https://doi.org/10.1007/s13277-014-2014-x.
- Chen, P., Zhang, J. Y., Sha, B. B., Ma, Y. E., Hu, T., Ma, Y. C., Sun, H., Shi, J.X., Dong, Z. M., Li, P. (2017). Luteolin inhibits cell proliferation and induces cell apoptosis via down-regulation of mitochondrial membrane potential in esophageal carcinoma cells EC1 and KYSE450. Oncotarget, 8(16), 27471. https://doi.org/10.18632/oncotarget.15832.
- Chen, J., Sun, M., Wang, X., Lu, J., Wei, Y., Tan, Y., Liu, Y., Götz, J., He, R., Hua, Q. (2014a). The herbal compound geniposide rescues formaldehyde-induced apoptosis in N2a neuroblastoma cells. Science China Life Sciences, 57, 412-421. https://doi.org/10.1007/s11427-014-4643-0.
- Demirci, S., Alp, C., Akşit, H., Ulutaş, Y., Altay, A., Yeniçeri, E., Köksal, E., Yaylı, N. (2023). Isolation, characterization and anticancer activity of secondary metabolites from Verbascum speciosum. Chemical Biology & Drug Design, 101(6), 1273-1282. https://doi.org/10.1111/cbdd.14211.
- Dinani, M. S., Malakooti, S., Akbari, V. (2020). In vitro cytotoxic activity of Verbascum alceoides against cervix carcinoma cells. Journal of Reports in Pharmaceutical Sciences, 9(1), 19.
https://doi.org/10.4103/jrptps.JRPTPS_65_19.
Ferino, A., Rapozzi, V., Xodo, L. E. (2020). The ROS-KRAS-Nrf2 axis in the control of the redox homeostasis and the intersection with survival-apoptosis pathways: Implications for photodynamic therapy. Journal of Photochemistry and Photobiology B: Biology, 202, 111672. https://doi.org/10.1016/j.jphotobiol.2019.111672.
- Gao, J., Fosbrook, C., Gibson, J., Underwood, T., Gray, J., Walters, Z. (2023). Targeting EZH2 in neuroblastoma. Cancer Treatment Reviews, 119, 102600. https://doi.org/10.1016/10.1016/j.ctrv.2023.102600.
- Garcia-Oliveira, P., Carreira-Casais, A., Pereira, E., Dias, M. I., Pereira, C., Calhelha, R. C., Stojkovic, D., Sokovic, M., Simal-Gandara, J., Prieto, M. A., Caleja, C. Barros, L. (2022). From tradition to health: Chemical and bioactive characterization of five traditional plants. Molecules, 27(19), 6495. https://doi.org/10.3390/molecules27196495.
- Gibellini, L., Pinti, M., Nasi, M., De Biasi, S., Roat, E., Bertoncelli, L., Cossarizza, A. (2010). Interfering with ROS metabolism in cancer cells: the potential role of quercetin. Cancers, 2(2), 1288–1311.
https://doi.org/10.3390/cancers2021288.
- Gourisankar, S., Krokhotin, A., Ji, W., Liu, X., Chang, C. Y., Kim, S. H., Li, Z., Wenderski, W., Simanauskaite, J. M., Yang, H., Vogel, H., Zhang, T., Green, M. R., Gray, N. S., Crabtree, G. R. (2023). Rewiring cancer drivers to activate apoptosis. Nature, 620(7973), 417-425.https://doi.org/10.1038/s41586-023-06348-2.
- Gökmen, A., Kúsz, N., Karaca, N., Demirci, F., Hohmann, J., Kırmızıbekmez, H. (2021). Secondary metabolites from Verbascum bugulifolium Lam. and their bioactivities. Natural Product Research, 35(23), 5294-5298. https://doi.org/10.1080/14786419.2020.1753052.
- Göse, M., Hacıoğlu, N. (2021). Bioactive compounds, antimicrobial and antibiofilm activity of two Verbascum species. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 24(3), 479-487. https://doi.org/10.18016/ksutarimdoga.v24i60916.750034.
- Green, D. R. (2022a). The mitochondrial pathway of apoptosis Part I: MOMP and beyond. Cold Spring Harbor Perspectives in Biology, 14(5), a041038. https://doi.org/10.1101/cshperspect.a041038.
- Green, D. R. (2022b). The mitochondrial pathway of apoptosis Part II: The BCL-2 protein family. Cold Spring Harbor Perspectives in Biology, 14(6), a041046. https://doi.org/10.1101/cshperspect.a041046.
- Hagemann, S., Misiak, D., Bell, J. L., Fuchs, T., Lederer, M. I., Bley, N., Hammerle, M., Ghazy, E., Sippl, W., Schulte, J. H., Hüttelmaier, S. (2023). IGF2BP1 induces neuroblastoma via a druggable feedforward loop with MYCN promoting 17q oncogene expression. Molecular Cancer, 22(1), 1-23. https://doi.org/10.1186/s12943-023-01792-0.
- Hu, X. Y., Liang, J. Y., Guo, X. J., Liu, L., Guo, Y. B. (2015). 5-Fluorouracil combined with apigenin enhances anticancer activity through mitochondrial membrane potential (DeltaPsim)-mediated apoptosis in hepatocellular carcinoma. Clinical and Experimental Pharmacology and Physiology, 42(2), 146–153. https://doi.org/10.1111/1440-1681.12333.
- Jacobson, J. C., Clark, R. A., Chung, D. H. (2023). High-risk neuroblastoma: A surgical perspective. Children, 10(2), 388. https://doi.org/10.3390/children10020388.
- Jeong, J. H., An, J. Y., Kwon, Y. T., Rhee, J. G., Lee, Y. J. (2009). Effects of low dose quercetin: cancer cell-specific inhibition of cell cycle progression. Journal of Cellular Biochemistry, 106(1), 73–82.
https://doi.org/10.1002/jcb.21977.
- Kachadourian, R., Day, B. J. (2006). Flavonoid-induced glutathione depletion: Potential implications for cancer treatment. Free Radical Biology and Medicine, 41(1), 65-76. https://doi.org/10.1016/j.freeradbiomed.2006.03.002.
- Keser, A. M., Yaprak, A. E. (2023). The assessment of genetic diversity and population structure of Verbascum gypsicola by ISSR markers for conservation purposes. Nordic Journal of Botany, 2023(2), e03660. https://doi.org/10.1111/njb.03660.
- Kızıltaş, H., Bingöl, Z., Gören, A. C., Alwasel, S. H., Gülçin, İ. (2022). Analysis of phenolic compounds by LC-HRMS and determination of antioxidant and enzyme inhibitory properties of Verbascum speciousum Schrad. Records of Natural Products, 17(3), 485-500. http://doi.org/10.25135/rnp.370.2210.2598.
- Klimek, B., Olszewska, M. A., Tokar, M. (2010). Simultaneous determination of flavonoids and phenylethanoids in the flowers of Verbascum densiflorum and V. phlomoides by high‐performance liquid chromatography. Phytochemical Analysis: An International Journal of Plant Chemical and Biochemical Techniques, 21(2), 150-156. http://doi.org/10.1002/pca.1171.
- Kopustinskiene, D. M., Jakstas, V., Savickas, A., Bernatoniene, J. (2020). Flavonoids as anticancer agents. Nutrients, 12(2), 457. http://doi.org/10.3390/nu12020457.
- Kovalevich, J., Langford, D. (2013). Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology. Neuronal Cell Culture: Methods and Protocols, 1078, 9-21.
http://doi.org/10.1007/978-1-62703-640-5_2.
- Lin, Y., Shi, R., Wang, X., Shen, H. M. (2008). Luteolin, a flavonoid with potential for cancer prevention and therapy. Current Cancer Drug Targets, 8(7), 634-646. http://doi.org/10.2174/156800908786241050.
- Lossi, L. (2022). The concept of intrinsic versus extrinsic apoptosis. Biochemical Journal, 479(3), 357-384. https://doi.org/10.1042/BCJ20210854.
- Lu, J., Papp, L. V., Fang, J., Rodriguez-Nieto, S., Zhivotovsky, B., Holmgren, A. (2006). Inhibition of mammalian thioredoxin reductase by some flavonoids: Implications for myricetin and quercetin anticancer activity. Cancer Research 66(8), 4410–4418. http://doi.org/10.2174/10.1158/0008-5472.CAN-05-3310.
- Luca, S. V., Miron, A., Aprotosoaie, A. C., Mihai, C. T., Vochita, G., Gherghel, D., Ciocarlan, N., Skalicka‐Woźniak, K. (2019). HPLC‐DAD‐ESI‐Q‐TOF‐MS/MS profiling of Verbascum ovalifolium Donn ex Sims and evaluation of its antioxidant and cytogenotoxic activities. Phytochemical Analysis, 30(1), 34-45. https://doi.org/10.1002/pca.2788.
- Mahmoud, S. M., Abdel-Azim, N. S., Shahat, A. A., Ismail, S. I., Hammouda, F. M. (2007). Phytochemical and biological studies on Verbascum sinaiticum growing in Egypt. Natural Product Sciences, 13(3), 186-189. https://doi.org/10.1001/s13003-133-186-89.
- Mellado, M., Madrid, A., Reyna, M., Weinstein-Oppenheimer, C., Mella, J., Salas, C. O., Sanchez, E., Cuellar, M. (2018). Synthesis of chalcones with antiproliferative activity on the SH-SY5Y neuroblastoma cell line: Quantitative structure–activity relationship models. Medicinal Chemistry Research, 27, 2414-2425. http://doi.org/10.1007/s00044-018-2245-2.
- Mihailović, V., Kreft, S., Benković, E. T., Ivanović, N., Stanković, M. S. (2016). Chemical profile, antioxidant activity and stability in stimulated gastrointestinal tract model system of three Verbascum species. Industrial Crops and Products, 89, 141-151. https://doi.org/10.1016/j.indcrop.2016.04.075.
- Morana, O., Wood, W., Gregory, C. D. (2022). The apoptosis paradox in cancer. International Journal of Molecular Sciences, 23(3), 1328. https://doi.org/10.3390/ijms23031328.
- Pk, N., Rajan, R. K., Nanchappan, V., Karuppaiah, A., Chandrasekaran, J., Jayaraman, S., Gunasekaran, V. (2023). C–Glucosyl Xanthone derivative Mangiferin downregulates the JNK3 mediated caspase activation in Almal induced neurotoxicity in differentiated SHSY-5Y neuroblastoma cells. Toxicology Mechanisms and Methods, 33(9), 707-718.
https://doi.org/10.1080/15376516.2023.2237106.
Pourmoslemi, S., Larki-Harchegani, A., Daneshyar, S., Dastan, D., Nili-Ahmadabadi, A., Jazaeri, M. (2023). Antibacterial and anti-glucosyltransferase activity of Verbascum speciosum against cariogenic streptococci. Journal of Pharmacopuncture, 26(2), 139. http://doi.org/10.3831/KPI.2023.26.2.139.
- Rahman, M. A., Yang, H., Lim, S. S., Huh, S. O. (2013). Apoptotic effects of Melandryum firmum root extracts in human SH-SY5Y neuroblastoma cells. Experimental Neurobiology, 22(3), 208.
http://doi.org/10.3831/10.5607/en.2013.22.3.208.
- Richeux, F., Cascante, M., Ennamany, R., Saboureau, D., Creppy, E. E. (1999). Cytotoxicity and genotoxicity of capsaicin in human neuroblastoma cells SHSY-5Y. Archives of Toxicology, 73, 403-409.
http://doi.org/10.1007/s002040050680.
- Selseleh, M., Ebrahimi, S. N., Aliahmadi, A., Sonboli, A., Mirjalili, M. H. (2020). Metabolic profiling, antioxidant, and antibacterial activity of some Iranian Verbascum L. species. Industrial Crops and Products, 153, 112609. https://doi.org/10.1016/j.indcrop.2020.112609.
- Shakeri, A. R., Farokh, A. (2015). Phytochemical evaluation and antioxidant activity of Verbascum sublobatum Murb. leaves. Research Journal of Pharmacognosy, 2(3), 43-47.
https://doi.org/10.1002/s20003-203-43-47.
- Shen, X., Si, Y., Wang, Z., Wang, J., Guo, Y., Zhang, X. (2016). Quercetin inhibits the growth of human gastric cancer stem cells by inducing mitochondrial-dependent apoptosis through the inhibition of PI3K/Akt signaling. International Journal of Molecular Medicine, 38(2), 619-626. http://doi.org/10.3892/ijmm.2016.2625.
- Shi, M. D., Shiao, C. K., Lee, Y. C., Shih, Y. W. (2015). Apigenin, a dietary flavonoid, inhibits proliferation of human bladder cancer T-24 cells via blocking cell cycle progression and inducing apoptosis. Cancer Cell ınternational, 15, 33. http://doi.org/10.1186/s12935-015-0186-0.
- Singh, P., Lim, B. (2022). Targeting apoptosis in cancer. Current Oncology Reports, 24(3), 273-284. http://doi.org/10.1007/s11912-022-01199-y.
- Singh, V., Khurana, A., Navik, U., Allawadhi, P., Bharani, K. K., Weiskirchen, R. (2022). Apoptosis and pharmacological therapies for targeting thereof for cancer therapeutics. Sci, 4(2), 15.
https://doi.org/10.3390/sci4020015.
- Wan, Y., Yang, L., Jiang, S., Qian, D., Duan, J. (2022). Excessive apoptosis in ulcerative colitis: crosstalk between apoptosis, ROS, ER stress, and intestinal homeostasis. Inflammatory bowel diseases, 28(4), 639-648. https://doi.org/10.1093/ibd/izab277.
- Slika, H., Mansour, H., Wehbe, N., Nasser, S. A., Iratni, R., Nasrallah, G., Shaito, A., Ghaddar, T., Koneissy, F., Eid, A. H. (2022). Therapeutic potential of flavonoids in cancer: ROS-mediated mechanisms. Biomedicine & Pharmacotherapy, 146, 112442. http://doi.org/10.1016/j.biopha.2021.112442.
- Souza, R. P., Bonfim-Mendonça, P. D. S., Gimenes, F., Ratti, B. A., Kaplum, V., Bruschi, M. L., Nakamura, C. V., Silva, S. O., Maria-Engler, S. S., Consolaro, M. E. (2017). Oxidative stress triggered by apigenin induces apoptosis in a comprehensive panel of human cervical cancer-derived cell lines. Oxidative Medicine and Cellular Longevity, 2017, 152745. http://doi.org/10.1155/2017/1512745.
- Srivastava, S., Somasagara, R. R., Hegde, M., Nishana, M., Tadi, S. K., Srivastava, M., Choudhary, B., Raghavan, S. C. (2016). Quercetin, a natural flavonoid interacts with DNA, arrests cell cycle and causes tumor regression by activating mitochondrial pathway of apoptosis. Scientific Reports, 6(1), 1-13. http://doi.org/10.1038/srep24049.
- Tai, K. K., Pham, L., & Truong, D. D. (2011). Idebenone induces apoptotic cell death in the human dopaminergic neuroblastoma SHSY-5Y cells. Neurotoxicity Research, 20, 321-328.
http://doi.org/10.1007/s12640-011-9245-z.
- Talib, W. H., & Mahasneh, A. M. (2010a). Antiproliferative activity of plant extracts used against cancer in traditional medicine. Scientia Pharmaceutica, 78(1), 33-46.
http://doi.org/10.3797/scipharm.0912-11.
- Talib, W. H., & Mahasneh, A. M. (2010b). Antimicrobial, cytotoxicity and phytochemical screening of Jordanian plants used in traditional medicine. Molecules, 15(3), 1811-1824.
http://doi.org/10.3390/molecules15031811.
- Taşkaya, A., Şahin, B., Güvensen, N. C., Mammadov, R. (2023). A preliminary dtudy on anticancer and antimicrobial potential of methanolic extracts of Verbascum napifolium. ISPEC Journal of Agricultural Sciences, 7(1), 135-145. https://doi.org/10.5281/zenodo.7749361.
- Tatli, I. I., Akdemir, Z. F. (2006). Traditional uses and biological activities of Verbascum species. FABAD Journal of Pharmaceutical Sciences, 31(2), 85-96. https://doi.org/10.1003/s31002-312-185-96.
- Tatli, I. I., Akdemir, Z. S., Yesilada, E., & Küpeli, E. (2008). Anti-inflammatory and antinociceptive potential of major phenolics from Verbascum salviifolium Boiss. Zeitschrift für Naturforschung C, 63(3-4), 196-202. https://doi.org/10.1515/znc-2008-3-406.
- Teekaraman, D., Elayapillai, S. P., Viswanathan, M. P., Jagadeesan, A. (2019). Quercetin inhibits human metastatic ovarian cancer cell growth and modulates components of the intrinsic apoptotic pathway in PA-1 cell line. Chemico-Biological Interactions, 300, 91-100. https://doi.org/10.1016/j.cbi.2019.01.008.
- Vetter, I., Mozar, C. A., Durek, T., Wingerd, J. S., Alewood, P. F., Christie, M. J., Lewis, R. J. (2012). Characterisation of Nav types endogenously expressed in human SH-SY5Y neuroblastoma cells. Biochemical Pharmacology, 83(11), 1562-1571. https://doi.org/10.1016/j.bcp.2012.02.022.
- Wang, B., Zhao, X. H. (2017). Apigenin induces both intrinsic and extrinsic pathways of apoptosis in human colon carcinoma HCT-116 cells. Oncology Reports, 37(2), 1132-1140.https://doi.org/10.3892/or.2016.5303.
- Zhao, Y. L., Wang, S. F., Li, Y., He, Q. X., Liu, K. C., Yang, Y. P., & Li, X. L. (2011). Isolation of chemical constituents from the aerial parts of Verbascum thapsus and their antiangiogenic and antiproliferative activities. Archives of Pharmacal Research, 34, 703-707. https://doi.org/10.1007/s12272-011-0501-9.
- Zhou, M., Shen, S., Zhao, X., Gong, X. (2017). Luteoloside induces G0/G1 arrest and prodeath autophagy through the ROS-mediated AKT/mTOR/p70S6K signalling pathway in human non-small cell lung cancer cell lines. Biochemical and Biophysical Research Communications, 94(1–2), 263–269. https://doi.org/10.1016/j.bbrc.2017.10.042.