Sains Malaysiana 52(12)(2023): 3533-3550

http://doi.org/10.17576/jsm-2023-5212-15

Pyranocycloartobiloxanthone A Suppressed Metastasis Ovarian Cancer Cells via S Phase Cell Cycle Arrest and Apoptosis

(Piranosikloartobilozanton A Menindas Metastasis Sel Kanser Ovari melalui Hentian Kitaran Sel Fasa S dan Apoptosis)

MASHITOH ABD RAHMAN¹, NAJIHAH MOHD HASHIM^1,2,* & IDRIS ADEWALE AHMED³

¹Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universiti Malaya, 50603 Kuala Lumpur, Malaysia

²Centre for Natural Products Research and Drug Discovery (CENAR), Universiti Malaya, 50603 Kuala Lumpur, Malaysia

³Department of Biotechnology, Faculty of Applied Science, Lincoln University, Kelana Jaya 47301 Petaling Jaya, Selangor, Malaysia

Diserahkan: 14 April 2023/Diterima: 11 Disember 2023

Abstract

Ovarian cancer is a deadly disease with a poor prognosis, highlighting the urgent need for novel therapeutic alternatives. Pyranocycloartobiloxanthone A (PA), an exceptional xanthone compound has garnered attention due to its diverse medicinal properties. This study aimed to investigate the anticancer properties of PA on metastatic ovarian cancer SKOV-3 cells. Cytotoxicity was evaluated using an MTT assay, while apoptotic mechanisms were determined using AO/PI double staining, annexin V-fluorescein isothiocyanate, multiple cytotoxicity-3, reactive oxygen species (ROS) production, caspases, real-time PCR, western blot, human apoptosis protein profile array and cell cycle analysis. PA inhibited SKOV-3 cell proliferation in a time-dependent manner, with an IC₅₀ of 7.0 ± 0.5 µg/mL and a selectivity index of 13.2 after 72 h of treatment. PA induced apoptosis through the intrinsic apoptotic pathway and arrested the cell cycle at the S phase. PA stimulated ROS production and disrupted the mitochondrial membrane potential, releasing cytochrome c from mitochondria to the cytosol. Additionally, results from human apoptotic protein profile indicated that 21 proteins were upregulated while 22 proteins were downregulated, including Bcl-2, survivin, and HSP70. These findings suggest that PA has the potential as a lead molecule in the development of a chemotherapy drug for ovarian cancer. However, further research is necessary to evaluate the safety and efficacy of PA in preclinical and clinical settings.

Keywords: Apoptosis; Artocarpus obtusus; cell cycle; ovarian cancer; pyranocycloartobiloxanthone A

Abstrak

Kanser ovari adalah penyakit yang membawa maut dengan prognosis yang kurang baik, menunjukkan keperluan yang mendesak untuk terapi alternatif yang novel. Piranosikloartobilozanton A (PA), sebatian zanton yang luar biasa telah menarik perhatian kerana sifat perubatannya yang pelbagai. Penyelidikan ini bertujuan untuk mengkaji sifat antikanser PA ke atas sel kanser ovari metastatis SKOV-3. Sitotoksisiti dinilai menggunakan asai MTT, manakala mekanisme apoptosis ditentukan menggunakan pewarnaan berganda AO/PI, annexin V-fluoresein isotiosianat, pelbagai sitotoksisiti-3, pengeluaran spesies oksigen reaktif (ROS), caspases, masa nyata PCR, pemblotan western, profil protein apoptosis manusia dan analisis kitaran sel. PA merencat proliferasi sel SKOV-3 bergantung kepada masa dengan IC₅₀ 7.0 ± 0.5 µg/mL dan indeks selektiviti sebanyak 13.2 selepas 72 jam rawatan. PA aruhan apoptosis melalui laluan apoptosis intrinsik dan menahan kitaran sel pada fasa S. PA merangsang pengeluaran ROS dan mengganggu potensi membran mitokondria, melepaskan sitokrom c daripada mitokondria ke sitosol. Selain itu, hasil daripada profil protein apoptosis manusia menunjukkan bahawa pengawalaturan 21 protein telah meningkat manakala 22 protein telah menurun termasuk Bcl-2, survivin dan HSP70. Penemuan ini mencadangkan bahawa PA mempunyai potensi sebagai molekul utama dalam membangunkan dadah kemoterapi untuk kanser ovari. Walau bagaimanapun, kajian lanjut adalah perlu untuk menilai keselamatan dan keberkesanan PA dalam konteks praklinikal dan klinikal.

Kata kunci: Apoptosis; Artocarpus obtusus; kanser ovari; kitaran sel; piranosikloartobilozanton A

RUJUKAN

Albakova, Z., Armeev, G.A., Kanevskiy, L.M., Kovalenko, E.I. & Sapozhnikov, A.M. 2020. HSP70 multi-functionality in cancer. Cells 9(3): 587.

Albasher, G., AlKahtane, A.A., Alarifi, S., Ali, D., Alessia, M.S., Almeer, R.S., Abdel-Daim, M.M., Al-Sultan, N.K., Al-Qahtani, A.A., Ali, H. & Alkahtani, S. 2019. Methotrexate-induced apoptosis in human ovarian adenocarcinoma Skov-3 cells via ros-mediated Bax/Bcl-2-cyt-c release cascading. OncoTargets and Therapy 12: 21-30.

Braga, L.D.C., Gonçales, N.G., Furtado, R.D.S., Andrade, W.P.D., Silva, L.M. & Silva Filho, A.L.D. 2020. Apoptosis-related gene expression can predict the response of ovarian cancer cell lines to treatment with recombinant human trail alone or combined with cisplatin. Clinics 75: e1492.1-7.

Calderón-Montaño, J.M., Martínez-Sánchez, S.M., Burgos-Morón, E., Guillén-Mancina, E., Jiménez-Alonso, J.J., García, F., Aparicio, A. & López-Lázaro, M. 2019. Screening for selective anticancer activity of plants from Grazalema Natural Park, Spain. Natural Product Research 33(23): 3454-3458.

Chen, D., Li, B., Lei, T., Na, D., Nie, M., Yang, Y., Xie, C., He, Z. & Wang, J. 2021. Selective mediation of ovarian cancer SKOV3 cells death by pristine carbon quantum Dots/Cu₂O composite through targeting matrix metalloproteinases, angiogenic cytokines and cytoskeleton. Journal of Nanobiotechnology 19: 68.

Czabotar, P.E. & Garcia-Saez, A.J. 2023. Mechanisms of BCL-2 family proteins in mitochondrial apoptosis. Nature Reviews Molecular Cell Biology 24(10): 732-748.

Ditano, J.P., Sakurikar, N. & Eastman, A. 2021. Activation of CDC25A phosphatase is limited by CDK2/Cyclin A-mediated feedback inhibition. Cell Cycle 20(13): 1308-1319.

Gielecińska, A., Kciuk, M., Mujwar, S., Celik, I., Kołat, D., Kałuzińska-Kołat, Ż. & Kontek, R. 2023. Substances of natural origin in medicine: Plants vs. cancer. Cells 12(7): 986.

Green, D.R. 2022. Caspase activation and inhibition. Cold Spring Harbor Perspectives in Biology 14(8): a041020.

Hashemi, S.S., Amini-Farsani, Z., Rahmati, S., Jazaeri, A., Mohammadi-Samani, M. & Asgharzade, S. 2021. Oleuropein reduces cisplatin resistance in ovarian cancer by targeting apoptotic pathway regulators. Life Sciences 278: 119525.

Hashim, N.M., Rahmani, M., Ee, G.C.L., Sukari, M.A., Yahayu, M., Oktima, W., Ali, A.M. & Go, R. 2012. Antiproliferative activity of xanthones isolated from Artocarpus obtusus. Journal of Biomedicine and Biotechnology 2012: 130627.

Hashim, N.M., Rahmani, M., Sukari, M.A., Ali, A.M., Alitheen, N.B., Go, R. & Ismail, H.B.M. 2010. Two new xanthones from Artocarpus obtusus. Journal of Asian Natural Products Research12(2): 106-112.

Ivanova, V., Dikov, T. & Dimitrova, N. 2017. Histologic subtypes of ovarian carcinoma: Selected diagnostic and classification problems in Bulgaria: Is low hospital volume an issue? Tumori 103(2): 148-154.

Joly, F., Fabbro, M., Follana, P., Lequesne, J., Medioni, J., Lesoin, A., Frenel, J-S., Abadie-Lacourtoisie, S., Floquet, A., Gladieff, L., You, B., Gavoille, C., Kalbacher, E., Briand, M., Brachet, P-E., Giffard, F., Weiswald, L-B., Just, P-A., Blanc-Fournier, C., Leconte, A., Clarisse, B., Leary, A. & Poulain, L. 2022. A phase II study of Navitoclax (ABT-263) as single agent in women heavily pretreated for recurrent epithelial ovarian cancer: The MONAVI – GINECO study. Gynecologic Oncology 165(1): 30-39.

Kim, I.H., Eom, T., Park, J.Y., Kim, H.J. & Nam, T.J. 2022. Dichloromethane fractions of Calystegia soldanella induce S-phase arrest and apoptosis in HT-29 human colorectal cancer cells. Molecular Medicine Reports 25(2): 60.

Kitazumi, I. & Tsukahara, M. 2011. Regulation of DNA fragmentation: The role of caspases and phosphorylation. The FEBS Journal 278(3): 427-441.

Kong, X.X., Jiang, S., Liu, T., Liu, G.F. & Dong, M. 2020. Paclitaxel increases sensitivity of SKOV3 cells to hyperthermia by inhibiting heat shock protein 27. Biomedicine and Pharmacotherapy 132: 110907.

Lang, F., Qu, J., Yin, H., Li, L., Zhi, Y., Liu, Y., Fang, Z. & Hao, E. 2018. Apoptotic cell death induced by Z-ligustilidein human ovarian cancer cells and role of NRF2. Food and Chemical Toxicology 121: 631-638.

Lathiff, S.M., Jemaon, N., Abdullah, S.A. & Jamil, S. 2015. Flavonoids from Artocarpus anisophyllus and their bioactivities. Natural Products Communication 10(3): 393-396.

Li, J., Shi, J., Medina, J.E., Zhou, J., Du, X., Wang, H., Yang, C., Liu, J., Yang, Z., Dinulescu, D.M. & Xu, B. 2017. Selectively inducing cancer cell death by intracellular enzyme-instructed self-assembly (EISA) of dipeptide derivatives. Advanced Healthcare Materials 6(15): 1601400.

Li, Q., Tan, Q., Ma, Y., Gu, Z. & Chen, S. 2022. Myricetin suppresses ovarian cancer in vitro by activating the P38/Sapla signaling pathway and suppressing intracellular oxidative stress. Frontiers in Oncology 12: 903394.

López-Lázaro, M. 2015. How many times should we screen a chemical library to discover an anticancer drug? Drug Discovery Today 2(20): 167-169.

Maleki, N., Yavari, N., Ebrahimi, M., Faisal Faiz, A., Khosh Ravesh, R., Sharbati, A., Panji, M., Lorian, K., Gravand, A., Abbasi, M., Abazari, O., Shafiee Mehr, M. & Eskandari, Y. 2022. Silibinin exerts anti-cancer activity on human ovarian cancer cells by increasing apoptosis and inhibiting epithelial-mesenchymal transition (EMT). Gene 823: 146275.  

Marchetti, C., De Felice, F., Romito, A., Iacobelli, V., Sassu, C.M., Corrado, G., Ricci, C., Scambia, G. & Fagotti, A. 2021. Chemotherapy resistance in epithelial ovarian cancer: Mechanisms and emerging treatments. Seminars in Cancer Biology 77: 144-166.

Mohan, S., Abdelwahab, S.I., Kamalidehghan, B., Syam, S., May, K.S., Harmal, N.S.M., Shafifiyaz, N., Hadi, A.H.A., Hashim, N.M. & Rahmani, M. 2012. Involvement of NF-ΚB and Bcl2/Bax signaling pathways in the apoptosis of MCF7 cells induced by a xanthone compound Pyranocycloartobiloxanthone A. Phytomedicine 19(11): 1007-1015.

Nayak, M., Nagarajan, A., Majeed, M., Nagabhushanam, K. & Choudhury, A.K. 2018. In vitro anti-acne activity of phytoactives from the stem bark of Artocarpus hirsutus Lam. and characterisation of pyranocycloartobiloxanthone A as a mixture of two anomers. Natural Product Research 32(17): 2116-2120.

Pal, S., Bhowmick, S., Sharma, A., Sierra-Fonseca, J.A., Mondal, S., Afolabi, F. & Roy, D. 2023. Lymphatic vasculature in ovarian cancer. Biochimica et Biophysica Acta (BBA) - Reviews on Cancer 1878(5): 188950.

Pellegrini, E., Multari, G., Gallo, R.R., Vecchiotti, D., Zazzeroni, F., Condello, M. & Meschini, S. 2022. A natural product, voacamine, sensitizes paclitaxel-resistant human ovarian cancer cells. Toxicology and Applied Pharmacology 434: 115816.

Rahman, M.A., Ramli, F., Karimian, H., Dehghan, F., Nordin, N., Ali, H.M., Mohan, S. & Hashim, N.M. 2016. Artonin E induces apoptosis via mitochondrial dysregulation in SKOV-3 ovarian cancer cells. PLoS ONE 11(3): e0151466.

Rahman, M.A. & Hashim, N.M. 2022. Apoptotic induction mechanism of artonin E in 3D ovarian cancer cell lines. Indonesian Journal of Pharmacy 33(1): 147-158.

Rasouli, M., Mehdian, H., Hajisharifi, K., Amini, E., Ostrikov, K. & Robert, E. 2021. Plasma‐activated medium induces apoptosis in chemotherapy‐resistant ovarian cancer cells: High selectivity and synergy with carboplatin. Plasma Processes and Polymers 18(9): 2100074.

Schoutrop, E., Moyano-Galceran, L., Lheureux, S., Mattsson, J., Lehti, K., Dahlstrand, H. & Magalhaes, I. 2022. Molecular, cellular, and systemic aspects of epithelial ovarian cancer and its tumor microenvironment. Seminars in Cancer Biology 86: 207-223.

Sidahmed, H.M.A., Hashim, N.M., Amir, J., Abdulla, M.A., Hadi, A.H.A., Abdelwahab, S.I., Taha, M.M.E., Hassandarvish, P., Teh, X., Loke, M.F., Vadivelu, J., Rahmani, M. & Mohan, S. 2013. Pyranocycloartobiloxanthone A, a novel gastroprotective compound from Artocarpus obtusus Jarret, against ethanol-induced acute gastric ulcer in vivo. Phytomedicine 20(10): 834-843.

Singh, P. & Lim, B. 2022. Targeting apoptosis in cancer. Current Oncology Reports 24(3): 273-284.

Smirnova, I., Drăghici, G., Kazakova, O., Vlaia, L., Avram, S., Mioc, A., Mioc, M., Macaşoi, I., Dehelean, C., Voicu, A. & Şoica, C. 2022. Hollongdione arylidene derivatives induce antiproliferative activity against melanoma and breast cancer through pro-apoptotic and antiangiogenic mechanisms. Bioorganic Chemistry 119: 105535.

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.

Wang, Z., Guo, B., Yue, S., Zhao, S., Meng, F. & Zhong, Z. 2022. HER-2-mediated nano-delivery of molecular targeted drug potently suppresses orthotopic epithelial ovarian cancer and metastasis. International Journal of Pharmaceutics 625: 122126.

Wheatley, S.P. & Altieri, D.C. 2019. Survivin at a glance. Journal of Cell Science 132(7): p.jcs223826.

Wong, M.L. & Medrano, J.F. 2005. Real-time PCR for mRNA quantitation. BioTechniques 39(1): 75-85.

Worsley, C.M., Veale, R.B. & Mayne, E.S. 2022. Inducing apoptosis using chemical treatment and acidic pH and detecting it using the annexin V flow cytometric assay. PLoS ONE 17(6): e0270599.

Wu, G., Peng, H., Tang, M., Yang, M., Wang, J., Hu, Y., Li, Z., Li, J., Li, Z. & Song, L. 2021. ZNF711 down-regulation promotes cisplatin resistance in epithelial ovarian cancer via interacting with JHDM2A and suppressing SLC31A1 expression. EBioMedicine 71: 103558.

Yeung, T.L., Leung, C.S., Yip, K.P., Au Yeung, C.L., Wong, S.T. & Mok, S.C. 2015. Cellular and molecular processes in ovarian cancer metastasis. American Journal of Physiology - Cell Physiology 309(7): C444-C456.

Zhang, H., Gong, W., Wu, S. & Perrett, S. 2022. Hsp70 in redox homeostasis. Cells 11(5): 829.

*Pengarang untuk surat-menyurat; email: najihahmh@um.edu.my