Cytotoxicity and potential anticancer activities of herbal formulations from cultivated Ganoderma species in Nigeria against Human Hepatocellular Carcinoma
Resumo
Domesticated wild Ganoderma species have emerged as a valuable adjunct in the treatment of specific cancers and tumors, offering new avenues for therapeutic intervention. However, many existing treatments for Human Hepatocellular Carcinoma (HepG2), which are mostly costly, are limited by cirrhosis. In contrast, natural compounds in Ganoderma have potential synergistic and low-toxicity properties that could enhance therapeutic outcomes. In this study, heavy metal analysis, anti-proliferation screening of extracts and formulations of three cultivated Ganoderma species against HepG2 and Vero cells, and cytoplasmic fluorescence imaging of cells tracked with Red CMTPX were conducted. Ganoderma isolates were identified using Polymerase chain reactions (PCR) with Internal transcribed spacer sequences (ITS1 and ITS4) using a GenBank BLAST. Heavy metals were at acceptable levels according to the WHO standard. Indeed, the effective anti-proliferative, cell-mediated cytotoxicity of these species was observed. The hot water extract of Ganoderma sessile, Ganoderma oregonense, and Ganoderma mbrekobenum combined formulation (MCF-11) exhibited potent cytotoxicity with an IC50 of 44.7 µg mL-1 against HEPG2 cells, while showing mild toxicity with IC50 of 158.4 µg mL-1 against Vero cells. Mycelia biomass formulation of G. mbrekobenum showed a remarkable value of IC50 < 30 µg mL-1. At 100 μg mL-1, the effect of MCF-11 on treated cells was evident, as the cells lost their typical morphology and the cytoplasm appeared shrunken and rounded, with reduced cell volume. However, extracts demonstrated dose-dependent toxicity, indicating anti-HepG2 potential.
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Al-Qubaisi, M., Rozita, R., Yeap, S. K., Omar, A. R., Ali, A. M., & Alitheen, N. B. (2011). Selective cytotoxicity of goniothalamin against hepatoblastoma HepG2 cells. Molecules, 16(4), 2944-2959. https://doi.org/10.3390/molecules16042944
Anike, F. N., Isikhuemhen, O. S., Blum, D., & Neda, H. (2015). Nutrient requirements and fermentation conditions for mycelia and crude exo-polysaccharides production by Lentinus squarrosulus. Advances in Bioscience and Biotechnology, 6(8), 526-536. http://doi.org/10.4236/abb.2015.68055
Artun, F. T., Karagoz, A., Ozcan, G., Melikoglu, G., Anil, S., Kultur, S., & Sutlupinar, N. (2017). In vitro anticancer and cytotoxic activities of some plant extracts on HeLa and Vero cell lines. Proceedings, 1(10). https://doi.org/10.3390/proceedings1101019
Bray, F., Ferlay, J., Soerjomataram, I., Siegel, R. L., Torre, L. A., & Jemal., A. (2018). Global cancer statistics: GLOBOCAN estimates incidence and mortality worldwide for 36 cancers in 185 countries. Cancer Journal Clinicians, 68(6), 394–424. https://doi.org/10.3322/caac.21492
Brown, J. S., Amend, S. R., Austin, R. H., Gatenby, R. A., Hammarlund, E. U., & Pienta, K. J. (2023). Updating the definition of cancer. Molecular Cancer Research, 21(11), 1142-1147. https://doi.org/10.1158/1541-7786.MCR-23-0411
Chiu, H. I., Samad, N. A., Fang, L., & Lim, V. (2021). Cytotoxicity of targeted PLGA nanoparticles: A systematic review. RSC Advances, 11(16), 9433-9449. https://doi.org/10.1039/D1RA00074H
Demková, L., Árvay, J., Hauptvogl, M., Michalková, J., Šnirc, M., Harangozo, Ľ., Bobuľská, L., Bajčan, D., & Kunca, V. (2021). Mercury content in three edible wild-growing mushroom species from different environmentally loaded areas in Slovakia: An ecological and human health risk assessment. Journal of Fungi, 7(6). https://doi.org/10.3390/jof7060434
Dilna, D., Vidya, S. K., & Raj, B. M. (2014). Uptake of certain heavy metals from contaminated soil by mushroom-Galerina vittiformis. Ecotoxicology and Environmental Safety, 104, 414–422. https://doi.org/10.1016/j.ecoenv.2013.10.033
El-Serag, H. B. (2012). Epidemiology of viral hepatitis and hepatocellular carcinoma. Gastroenterology, 142(6), 1264–1273. https://doi.org/10.1053/j.gastro.2011.12.061
Forner, A., Reig, M., & Bruix, J. (2018). Hepatocellular carcinoma. The Lancet, 391(10127), 1301–1314. https://doi.org/10.1016/S0140-6736(18)30010-2
Gaobotse, G., Venkataraman, S., Brown, P. D., Masisi, K., Kwape, T. E., Nkwe, D. O., Rantong, G., & Makhzoum, A. (2023). The use of African medicinal plants in cancer management. Frontier Pharmacology 14. https://doi.org/10.3389/fphar.2023.1122388
Gill, S. K., & Rieder, M. J. (2008). Toxicity of a traditional Chinese medicine, Ganoderma lucidum, in children with cancer. Canadian Journal of Clinical Pharmacology, 15(2), e275-e285.
Huang, Y., Huang, S., Wei, W., Wu, Y., Jia, L., Du, Y., Luo, P. & Pan, W. (2024). Design, synthesis, and anticancer evaluation of novel half-sandwich Ru (II) complexes bearing pyrazalone moiety: Apoptosis inducers based on mitochondrial dysfunction and G0/G1 arrest, Journal of Inorganic Biochemistry, 250. https://doi.org/10.1016/j.jinorgbio.2023.112421
Hui, C. K., Majid, N. I., Zainol, M. K. M., Mohamad, H., & Zin, Z. M. (2018). Preliminary phytochemical screening and effect of hot water extraction conditions on phenolic contents and antioxidant capacities of Morinda citrifolia leaf. Malaysian Applied Biology, 47(4), 13–24.
Isikhuemhen, O. S., Nerud, F., Moncalvo, J. M., & Vilgalys, R. (2000). Mating compatibility and phylogeography in Pleurotus tuberregium. Mycological Research, 104(6), 732-737. https://doi.org/10.1017/S0953756299001926
Khalil, M. I. M., Ibrahim, M. M., El-Gaaly, G. A., & Sultan, A. S. (2015). Trigonella foenum (fenugreek) induced apoptosis in hepatocellular carcinoma cell line, HepG2, mediated by upregulation of p53 and proliferating cell nuclear antigen. BioMed Research International, 2015, 1–11. https://doi.org/10.1155/2015/914645
Khani, R., Moudi, M., & Khojeh, V. (2017). Contamination level, distribution, and health risk assessment of heavy and toxic metallic and metalloid elements in a cultivated mushroom, Pleurotus florida (Mont) Singer. Environmental Science and Pollution Research, 24, 4699–4708. https://doi.org/10.1007/s11356-016-8222-8
Lalotra, P., Gupta, D., Yangdol, R., Sharma, Y. P., & Gupta, S. K. (2016). Bioaccumulation of heavy metals in the sporocarps of some wild mushrooms. Current Research in Environmental & Applied Mycology, 6(3),159–165. http://doi/10.5943/cream/6/3/2
Lam, C. S., Cheng, L. P., Zhou, L. M., Cheung, Y. T., & Zuo, Z. (2020). Herb-drug interactions between the medicinal mushrooms Lingzhi and Yunzhi and cytotoxic anticancer drugs: A systematic review. Chinese Medicine, 15. https://doi.org/10.1186/s13020-020-00356-4
Lee, S. B., & Taylor, J. W. (1990). Isolation of DNA from fungal mycelia and single spores. In M. A. Innis, D. H. Gelfand, J. J. Sninsky, & T. J. White (Eds.), PCR protocols: A guide to methods and applications (pp. 282–287). Academic Press. https://doi.org/10.1016/b978-0-12-372180-8.50038-x
Liu, Y., Gao, J., Guan, J., Qian, Z., Feng, K., & Li, S. (2009). Evaluation of anti-proliferative activities and action mechanisms of extracts from two species of Ganoderma on tumor cell lines. Journal of Agriculture and Food Chemistry, 57(8), 3087-3093. https://doi.org/10.1021/jf900011f
Loyd, A. L., Barnes, C. W., Held, B. W., Schink, M. J., Smith, M. E., Smith, J. A., & Blanchette, R. A. (2018b). Elucidating "lucidum": Distinguishing the diverse laccate Ganoderma species of the United States. PLoS One. 13(7), e0199738. http://doi.org/10.1371/journal.pone.0199738
Loyd, A. L., Richter, B. S., Jusino, M. A., Truong, C., Smith, M. E., Blanchette, R. A., & Smith, J. A. (2018a). Identifying the “Mushroom of Immortality”: Assessing the Ganoderma species composition in commercial reishi products. Frontiers in Microbiology, 9. https://doi.org/10.3389/fmicb.2018.01557
Nwachukwu, V. A., Udedi, S. C., Ezeonu, F. C., Orji, F. A., Ezeanyanaso, C. S., Brai, B. I. C., Shode, F. O., & Elemo, G. N. (2017). An analysis of food value and some selected secondary metabolites of Emilia coccinea (Asteraceae) leaf. Journal of Complementary and Alternative Medical Research, 2(2), 1–11. https://doi.org/10.9734/JOCAMR/2017/29763
Nwogu, V. K., Udenze, E. C., & Chinedu, N. S. (2022). Immunological impacts of Ganoderma lucidum in Nigerian traditional medicine. Journal of Ethnopharmacology, 284.
Ofodile, L. N., Abraham, A., Ayoade, Y., Adamu, G. L., Nwakanma, M. N. C., Ovioma, G. O., Bikomo, O. E., Ikegwu, E., & Ayodeji, A. A. (2020a). Effect of the aqueous extract of Ganoderma lucidum on the haematology, estradiol, cholesterol, and protein levels of Wistar rats fed with monosodium glutamate. Malaysian Journal of Pharmaceutical Sciences, 18(2), 47-62. https://doi.org/10.21315/mjps2020.18.2.4
Ofodile, L. N., Isikhuemhen, O. S., Anike, F. N., & Adekunle, A. A. (2022). Domestication and cultivation studies on indigenous Ganoderma species from Nigeria. International Journal of Medicinal Mushrooms, 24(6), 69-78. http://doi.org/10.1615/IntJMedMushrooms.2022043906
Ofodile, L. N., Nicholas-Okpara, Nwachukwu, V. A. Ani, E., Ikegwu, E. M., Saanu, A.I., Ezenwa, P. C., Osorinde, R. T. (2020b). Production and nutritional composition of juice powder from oyster mushroom Pleurotus ostreatus (Jacq.) Kummer. Functional Foods in Health and Disease, 10(11), 482-492. https://doi.org/10.31989/ffhd.v10i11.751
Ofodile, L., Bikomo, E., Nicholas-okpara, V., Adekunle, A., Ani, E., & Kanife, U. (2025). Antitumour activity of Ganoderma mbrekobenum on monosodium glutamate induced uterine tumour in Wistar rats (Rattus norvegicus). Bozok Journal of Science, 3(1), 20–33. https://doi.org/10.70500/bjs.1565734
Olou, B., Langer, E., Ryvarden, L., Krah, F. S., Hounwanou, G., Piepenbring, M., & Yorou, N. (2023). New records and barcode sequence data of wood-inhabiting polypores in Benin with notes on their phylogenetic placements and distribution. Fungal Systematics and Evolution 11, 11-42. https://doi.org/10.3114/fuse.2023.11.02
Otto, E. C., Paloi, S., Peterson, S. W., Phosri, C., Roux, J., Salazar, W. A., Sánchez, A., Sarria, G. A., Shin, H. D., Silva, B. D., Silva, G. A., Smith, M. T., Souza-Motta, C. M., Stchigel, A. M., Stoilova-Disheva, M. M., Sulzbacher, M. A., Telleria, M. T., Toapanta, C., Traba, J. M.,... Groenewald, J. Z. (2016). Fungal Planet description sheets: 400–468. Persoonia, 36, 316–458. https://doi.org/10.3767/003158516X692185
Owen, R. W., Giacosa, A., Hull, W. E., Haubner, R., Spiegelhalder, B., & Bartsch, H. (2000). The antioxidant/anticancer potential of phenolic compounds isolated from olive oil. European Journal of Cancer, 36(10), 1235–1247. https://doi.org/10.1016/s0959-8049(00)00103-9
Pocasap, P., Nonpunya, A., & Weerapreeyakul, N. (2021). Pinus kesiya Royle ex Gordon induces apoptotic cell death in hepatocellular carcinoma HepG2 cell via intrinsic pathway by PARP and Topoisomerase I suppression. Biomedicine & Pharmacotherapy, 139. https://doi.org/10.1016/j.biopha.2021.111628.
Rai, A., Rai, P. K., Singh, S., & Sharma, N. K. (2015). Environmental factors affecting edible and medicinal mushroom production. In Production Techniques of Tropical Mushrooms in India (1st ed., pp. 67–81). Nirmal Publisher.
Saba, M., Falandysz, J., & Nnorom, I. C. (2016). Mercury bioaccumulation by Suillus bovinus mushroom and probable dietary intake with the mushroom meal. Environmental Science and Pollution Research, 23, 14549–14559. https://doi.org/doi/10.1007/s11356-016-6558-8
Serrano-Márquez, L., Trigos, Á., Couttolenc, A., Padron, J. M., Shnyreva, A.V., Mendoza, G. (2021). Anti-proliferative and antibacterial activity of extracts of Ganoderma strains grown in vitro. Food Science and Biotechnology, 30, 711–721. https://doi.org/10.1007/s10068-021-00903-1
Seyfferth, A. L., McClatchy, C., Paukett, M. (2016). Arsenic, lead, and cadmium in U.S. mushrooms and substrate in relation to dietary exposure. Environmental Science & Technology, 50, 9661–9670. https://doi.org/10.1021/acs.est.6b02133
Singal, A. G., Lampertico, P., & Nahon, P. (2020). Epidemiology and surveillance for hepatocellular carcinoma: New trends. Journal of Hepatology, 72(2), 250–261. https://doi.org/10.1016/j.jhep.2019.08.025
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. https://doi.org/10.3322/caac.21660
Swetha, M., Keerthana, C. K., Rayginia, T. P., Nath, L. R., Haritha, N. H., Shabna, A., Kalimuthu, K., Thangarasu, A. K., Aiswarya, S. U., Jannet, S., Pillai, S., Harikumar, K. B., Sundaram, S., Anto, N. P., Wu, D. H., Lankalapalli, R. S., Towner, R., Isakov, N., Deepa, S. S., ... Anto, R. J. (2022). Augmented efficacy of Uttroside B over sorafenib in a murine model of human hepatocellular carcinoma. Pharmaceuticals, 15(5). https://doi.org/10.3390/ph15050636
Tavakoli, J., Miar, S., Zadehzare, M. M., & Akbari, H. (2012). Evaluation of effectiveness of herbal medication in cancer care: A review study. Iranian Journal of Cancer Prevention, 5(3), 144–156. https://pubmed.ncbi.nlm.nih.gov/25628834/
Venook, A. P., Papandreou, C., Furuse, J., & De Guevara, L. L. (2010). The incidence and epidemiology of hepatocellular carcinoma: A global and regional perspective. The Oncologist, 15(Suppl. 4), 5–13. https://doi.org/10.1634/theoncologist.2010-s4-05
World Health Organization. (2007). WHO guidelines for assessing quality of herbal medicines with reference to contaminants and residues. https://www.who.int/publications/i/item/9789241594448
Wu, P., Zhang, C., Yin, Y., Zhang, X., Li, Q., Yuan, L., Sun, Y., Zhou, S., Ying, S., & Wu, J. (2024). Bioactivities and industrial standardization status of Ganoderma lucidum: A comprehensive review. Heliyon, 10(19), e36987. https://doi.org/10.1016/j.heliyon.2024.e36987
Wuana, R. A., & Okieimen, F. E. (2011). Heavy metals in contaminated soils: A review of sources, chemistry, risks and best available strategies for remediation. International Scholarly Research Notices, 2011, Article 402647, 1–20. https://doi.org/10.5402/2011/402647.
Yang, J. D., Hainaut, P., Gores, G. J., Amadou, A., Plymoth, A., & Roberts, L. R. (2019). A global view of hepatocellular carcinoma: Trends, risk, prevention and management. Nature Reviews Gastroenterology & Hepatology, 16(10), 589–604. http://doi.org/10.1038/s41575-019-0186-y
Zhang, Q., Lin, L., & Ye, W. (2018) Techniques for extraction and isolation of natural products: a comprehensive review. Chinese Medicine, 13. https://doi.org/10.1186/s13020-018-0177-x
Copyright (c) 2025 Lauretta Nwanneka Ofodile, Viola Anuli Nicholas-Okpara, Leonard Williams, Oniovosa Leonard Adamu-Governor, Uche Claris Kanife, Emmanuel Ani, Doyinsola Akinjayeju, Utom-Obong Udom Akpan, Emmanuel Mmaduabuchi Ikegwu, Adekunle Ayo Ayodeji, Ndubuisi Moses Chikere Nwakanma, Ige Saanu Anjorin (Autor)

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