Abstract
The mycelial extract of the entomopathogenic fungus Ophiocordyceps sobolifera has recently been reported to have anti-bacterial, anti-fungal and anti-cancer effects; however, there have been no reports on the medicinal value of the crude protein against the Michigan Cancer Foundation-7 (MCF-7) human breast cancer cell line. Therefore, in this study, crude proteins from 11 isolates of O. sobolifera were subjected to screening and evaluation for potential anticancer activity against a breast cancer cell line. The cytotoxic and antiproliferative effects of the crude proteins on human breast cancer MCF-7 cell line was investigated using the sulforhodamine B (SRB) assay and colony formation. The gene expression and cell cycle-associated protein levels were assayed by reverse transcription-quantitative polymerase chain reaction and western blotting analysis. The results showed that the crude proteins of all isolates had anti-cancer activity in a time- and dose-dependent manner. Among them, isolates Cod-KK1524, Cod-KK1630, Cod-KK1643 and Cod-KS1601 had greater inhibitory effects against cancer cells; therefore, these were chosen for further evaluation of anti-cancer mechanisms. These crude proteins exhibited a potent anticancer effect on the MCF-7 cells by inhibiting colony formation in a dose-dependent manner. Furthermore, the crude proteins significantly decreased the cyclin-dependent kinase 6 (cdk6) and cyclin D1 expression. Additionally, crude proteins significantly enhanced the potent cyclin-dependent kinase inhibitor (p21) levels, and reduced the expression level of the cell cycle protein, cyclin D1. These findings indicate that the crude protein of the entomopathogenic fungus O. sobolifera may be useful for developing an anti-cancer drug candidate for breast cancer therapeutic treatment.
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References
Agarwal G, Pradeep PV, Aggarwal V, Yip CH, Cheung PS (2007) Spectrum of breast cancer in Asian women. World J Surg 31:1031–1040. https://doi.org/10.1007/s00268-005-0585-9
Akin S, Babacan T, Sarici F, Altundag K (2014) A novel targeted therapy in breast cancer: cyclin dependent kinase inhibitors. J BUON 19:42–46
Arnold A, Papanikolaou A (2005) Cyclin D1 in breast cancer pathogenesis. J Clin Oncol 23:4215–4224. https://doi.org/10.1200/JCO.2005.05.064
Bai KC, Sheu F (2018) A novel protein from edible fungi Cordyceps militaris that induces apoptosis. J Food Drug Anal 26:21–30. https://doi.org/10.1016/j.jfda.2016.10.013
Buranrat B, Senggunprai L, Prawan A, Kukongviriyapan V (2016) Simvastatin and atorvastatin as inhibitors of proliferation and inducers of apoptosis in human cholangiocarcinoma cells. Life Sci 153:41–49. https://doi.org/10.1016/j.lfs.2016.04.018
Buranrat B, Suwannaloet W, Naowaboot J (2017a) Simvastatin potentiates doxorubicin activity against MCF-7 breast cancer cells. Oncol Lett 14:6243–6250. https://doi.org/10.3892/ol.2017.6783
Buranrat B, Mairuae N, Konsue A (2017b) Cratoxy formosum leaf extract inhibits proliferation and migration of human breast cancer MCF-7 cell. Biomed Pharmacother 90:77–84. https://doi.org/10.1016/j.biopha.2017.03.032
Buranrat B, Sangdee K, Sangdee A (2019) Comparative study on the effect of aqueous and ethanolic mycelial extracts from Polycephalomyces nipponicus (Ascomycetes) against human breast cancer MCF-7 Cells. Int J Med Mushrooms 21(7):671–681. https://doi.org/10.1615/IntJMedMushrooms.2019031140
Courjal F, Louason G, Speiser P, Katsaros D, Zeillinger R, Theillet C (1996) Cyclin gene amplification and overexpression in breast and ovarian cancers: evidence for the selection of cyclin D1 in breast and cyclin E in ovarian tumors. Int J Cancer 69:247–253. https://doi.org/10.1002/(SICI)1097-0215(19960822)69:4<247::AID-IJC1>3.0.CO;2-X
Deslouches B, Di YP (2017) Antimicrobial peptides with selective antitumor mechanisms: prospect for anticancer applications. Oncotarget 8(28):46635–46651. https://doi.org/10.18632/oncotarget.16743
Ezhevsky SA, Ho A, Becker-Hapak M, Davis PK, Dowdy SF (2001) Differential regulation of retinoblastoma tumor suppressor protein by G1 cyclin-dependent kinase complexes in vivo. Mol Cell Biol 21:4773–4784. https://doi.org/10.1128/MCB.21.14.4773-4784.2001
Ghoncheh M, Momenimovahed Z, Salehiniya H (2016) Epidemiology, incidence and mortality of breast cancer in Asia. Asian Pac J Cancer Prev 17(S3):47–52. https://doi.org/10.7314/apjcp.2016.17.s3.47
Gillett C, Smith P, Gregory W, Richards M, Millis R, Peters G, Barnes D (1996) Cyclin D1 and prognosis in human breast cancer. Int J Cancer 69:92–99. https://doi.org/10.1002/(SICI)1097-0215(19960422)69:2<92::AID-IJC4>3.0.CO;2-Q
Gray-Bablin J, Zalvide J, Fox MP, Knickerbocker CJ, DeCaprio JA, Keyomarsi K (1996) Cyclin E, a redundant cyclin in breast cancer. Proc Natl Acad Sci USA 93:15215–15220. https://doi.org/10.1073/pnas.93.26.15215
Harper JW, Elledge SJ, Keyomarsi K, Dynlacht B, Tsai LH, Zhang P, Dobrowolski S, Bai C, Connell-Crowley L, Swindell E, Fox MP, Wei N (1995) Inhibition of cyclin-dependent kinases by p21. Mol Biol Cell 6:387–400. https://doi.org/10.1091/mbc.6.4.387
Huang L, Li Q, Chen Y, Wang X, Zhou X (2009) Determination and analysis of cordycepin and adenosine in the products of Cordyceps spp. Afr J Microbiol Res 3:957–961. http://www.academicjournals.org/ajmr
Jaihan P, Sangdee K, Sangdee A (2016) Selection of entomopathogenic fungus for biological control of chili anthracnose disease caused by Colletotrichum spp. Eur J Plant Pathol 146:551–564. https://doi.org/10.1007/s10658-016-0941-7
Lamb R, Lehn S, Rogerson L, Clarke RB, Landberg G (2013) Cell cycle regulators cyclin D1 and CDK4/6 have estrogen receptor-dependent divergent functions in breast cancer migration and stem cell-like activity. Cell Cycle 12:2384–2394. https://doi.org/10.4161/cc.25403
Lundberg AS, Weinberg RA (1998) Functional inactivation of the retinoblastoma protein requires sequential modification by at least two distinct cyclin-cdk complexes. Mol Cell Biol 18:753–761. https://doi.org/10.1128/mcb.18.2.753
McIntosh GG, Anderson JJ, Milton I, Steward M, Parr AH, Thomas MD, Henry JA, Angus B, Lennard TW, Horne CH (1995) Determination of the prognostic value of cyclin D1 overexpression in breast cancer. Oncogene 11:885–891. https://www.ncbi.nlm.nih.gov/pubmed/7675447
O’Driscoll L, Clynes M (2006) Biomarkers and multiple drug resistance in breast cancer. Curr Cancer Drug Targets 6:365–384. https://doi.org/10.2174/156800906777723958
Pagani O, Senkus E, Wood W, Colleoni M, Cufer T, Kyriakides S, Costa A, Winer EP, Cardoso F, ESO-MBC Task Force (2010) International guidelines for management of metastatic breast cancer: can metastatic breast cancer be cured? J Natl Cancer Inst 102:456–463. https://doi.org/10.1093/jnci/djq029
Park BT, Na KH, Jung EC, Park JW, Kim HH (2009) Antifungal and anticancer activities of a protein from the mushroom Cordyceps militaris. Korean J Physiol Pharmacol 3:49–54. https://doi.org/10.4196/kjpp.2009.13.1.49
Sangdee K, Nakbanpote W, Sangdee A (2015) Isolation of the entomopathogenic fungal strain Cod-MK1201 from a cicada nymph and assessment of its antibacterial activities. Int J Med Mushrooms 17:51–63. https://doi.org/10.1615/intjmedmushrooms.v17.i1.60
Sangdee K, Buranrat B, Jaihan P, Thongchai S, Sangdee A (2018a) Evaluation of antibacterial and anticancer activities of the medicinal fungus Ophiocordyceps sobolifera (Ascomycetes) from Thailand. Int J Med Mushrooms 20:471–484. https://doi.org/10.1615/IntJMedMushrooms.2018026247
Sangdee A, Sangdee K, Buranrat B, Thammawat S (2018b) Effects of mycelial extract and crude protein of the medicinal mushroom, Ophiocordyceps sobolifera, on the pathogenic fungus, Candida albicans. Trop J Pharm Res 17(12):2449–2454. https://doi.org/10.4314/tjpr.v17i12.21
Sherr CJ, Roberts JM (1995) Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev 9:1149–1163. https://doi.org/10.1101/gad.9.10.1149
Wang SX, Liu Y, Zhang GQ, Zhao S, Xu F, Geng XL, Wang HX (2012) Cordysobin, a novel alkaline serine protease with HIV-1 reverse transcriptase inhibitory activity from the medicinal mushroom Cordyceps sobolifera. J Biosci Bioeng 113:42–47. https://doi.org/10.1016/j.jbiosc.2011.09.005
Wang ST, Ho HJ, Lin JT, Shieh JJ, Wu CY (2017) Simvastatin-induced cell cycle arrest through inhibition of STAT3/SKP2 axis and activation of AMPK to promote p27 and p21 accumulation in hepatocellular carcinoma cells. Cell Death Dis 23(2):8. https://doi.org/10.1038/cddis.2016.472
Wong JH, Ng TB, Wang H, Sze SC, Zhang KY, Li Q, Lu X (2011) Cordymin, an antifungal peptide from the medicinal fungus Cordyceps militaris. Phytomedicine 18:387–392. https://doi.org/10.1016/j.phymed.2010.07.010
Wu JY, Chen CH, Chang WH, Chung KT, Liu YW, Lu FJ, Chen CH (2011) Anti-cancer effects of protein extracts from Calvatia lilacina, Pleurotus ostreatus and Volvariella volvacea. Evid-Based Complementary Altern Med. https://doi.org/10.1093/ecam/neq057
Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R, Beach DV (1993) p21 is a universal inhibitor of cyclin kinases. Nature 366:701–704. https://doi.org/10.1038/366701a0
Yang S, Zhang H (2016) Optimization of the fermentation process of Cordyceps sobolifera Se-CEPS and its anti-tumor activity in vivo. J Biol Eng 10:1–8. https://doi.org/10.1186/s13036-016-0029-0
Zhang H, Hannon J, Beach D (1994) p21-containing cyclin kinases exist in both active and inactive states. Genes Dev 8:1750–1758. https://doi.org/10.1101/gad.8.15.1750
Zhang N, Yang Y, Cheng L, Zhang XM, Zhang S, Wang W, Liu SY, Wang SY, Wang RB, Xu WJ, Dai L, Yan N, Fan P, Dai LX, Tian HW, Liu L, Deng HX (2012) Combination of Caspy2 and IP-10 gene therapy significantly improves therapeutic efficacy against murine malignant neoplasm growth and metastasis. Hum Gene Ther 23:837–846. https://doi.org/10.1089/hum.2011.136
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The authors express their thanks to Mahasarakham University Faculty of Science for providing financial support for this study and the authors gratefully thank to the Mahasarakham University Faculty of Medicine for providing equipment.
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The author Aphidech Sangdee designed, performed, analyzed the experiments and wrote the manuscript. Benjaporn Buranrat designed, performed and analyzed the experiments. Kusavadee Sangdee and Sutthiwan Thammawat critically proofed the data and manuscript.
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Buranrat, B., Sangdee, K., Thammawat, S. et al. Mechanisms of crude protein from medicinal mushroom Ophiocordyceps sobolifera against human breast MCF-7 cancer cells. Biologia 75, 1759–1768 (2020). https://doi.org/10.2478/s11756-020-00482-2
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DOI: https://doi.org/10.2478/s11756-020-00482-2