Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro
Background The highly valued medicinal tiger milk mushroom (also known as Lignosus rhinocerus) has the ability to cure numerous ailments. Its anticancer activities are well explored, and recently a partially purified cytotoxic protein fraction termed F5 from the mushroom’s sclerotial cold water extr...
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PeerJ Inc.
2018-06-01
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| author | Hui Yeng Y. Yap Nget Hong Tan Szu Ting Ng Chon Seng Tan Shin Yee Fung |
| author_facet | Hui Yeng Y. Yap Nget Hong Tan Szu Ting Ng Chon Seng Tan Shin Yee Fung |
| author_sort | Hui Yeng Y. Yap |
| collection | DOAJ |
| description | Background The highly valued medicinal tiger milk mushroom (also known as Lignosus rhinocerus) has the ability to cure numerous ailments. Its anticancer activities are well explored, and recently a partially purified cytotoxic protein fraction termed F5 from the mushroom’s sclerotial cold water extract consisting mainly of fungal serine proteases was found to exhibit potent selective cytotoxicity against a human breast adenocarcinoma cell line (MCF7) with IC50 value of 3.00 μg/ml. However, characterization of its cell death-inducing activity has yet to be established. Methods The mechanism involved in the cytotoxic activities of F5 against MCF7 cells was elucidated by flow cytometry-based apoptosis detection, caspases activity measurement, and expression profiling of apoptosis markers by western blotting. Molecular attributes of F5 were further mined from L. rhinocerus’s published genome and transcriptome for future exploration. Results and Discussion Apoptosis induction in MCF7 cells by F5 may involve a cross-talk between the extrinsic and intrinsic apoptotic pathways with upregulation of caspase-8 and -9 activities and a marked decrease of Bcl-2. On the other hand, the levels of pro-apoptotic Bax, BID, and cleaved BID were increased accompanied by observable actin cleavage. At gene level, F5 composed of three predicted non-synonymous single nucleotide polymorphisms (T > C) and an alternative 5′ splice site. Conclusions Findings from this study provide an advanced framework for further investigations on cancer therapeutics development from L. rhinocerus. |
| first_indexed | 2024-03-09T06:23:08Z |
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| institution | Directory Open Access Journal |
| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T06:23:08Z |
| publishDate | 2018-06-01 |
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| spelling | doaj.art-c38b3db3f1a840db8b3501c033cb57b42023-12-03T11:33:03ZengPeerJ Inc.PeerJ2167-83592018-06-016e494010.7717/peerj.4940Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitroHui Yeng Y. Yap0Nget Hong Tan1Szu Ting Ng2Chon Seng Tan3Shin Yee Fung4Department of Oral Biology, Faculty of Dentistry, MAHSA University, Kuala Lumpur, MalaysiaMedicinal Mushroom Research Group, Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Wilayah Persekutuan, MalaysiaLigno Biotech Sdn Bhd, Balakong Jaya, Selangor, MalaysiaLigno Biotech Sdn Bhd, Balakong Jaya, Selangor, MalaysiaMedicinal Mushroom Research Group, Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Wilayah Persekutuan, MalaysiaBackground The highly valued medicinal tiger milk mushroom (also known as Lignosus rhinocerus) has the ability to cure numerous ailments. Its anticancer activities are well explored, and recently a partially purified cytotoxic protein fraction termed F5 from the mushroom’s sclerotial cold water extract consisting mainly of fungal serine proteases was found to exhibit potent selective cytotoxicity against a human breast adenocarcinoma cell line (MCF7) with IC50 value of 3.00 μg/ml. However, characterization of its cell death-inducing activity has yet to be established. Methods The mechanism involved in the cytotoxic activities of F5 against MCF7 cells was elucidated by flow cytometry-based apoptosis detection, caspases activity measurement, and expression profiling of apoptosis markers by western blotting. Molecular attributes of F5 were further mined from L. rhinocerus’s published genome and transcriptome for future exploration. Results and Discussion Apoptosis induction in MCF7 cells by F5 may involve a cross-talk between the extrinsic and intrinsic apoptotic pathways with upregulation of caspase-8 and -9 activities and a marked decrease of Bcl-2. On the other hand, the levels of pro-apoptotic Bax, BID, and cleaved BID were increased accompanied by observable actin cleavage. At gene level, F5 composed of three predicted non-synonymous single nucleotide polymorphisms (T > C) and an alternative 5′ splice site. Conclusions Findings from this study provide an advanced framework for further investigations on cancer therapeutics development from L. rhinocerus.https://peerj.com/articles/4940.pdfF5Tiger milk mushroomSerine proteaseLignosus rhinocerusAnticancerApoptosis |
| spellingShingle | Hui Yeng Y. Yap Nget Hong Tan Szu Ting Ng Chon Seng Tan Shin Yee Fung Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro PeerJ F5 Tiger milk mushroom Serine protease Lignosus rhinocerus Anticancer Apoptosis |
| title | Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro |
| title_full | Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro |
| title_fullStr | Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro |
| title_full_unstemmed | Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro |
| title_short | Molecular attributes and apoptosis-inducing activities of a putative serine protease isolated from Tiger Milk mushroom (Lignosus rhinocerus) sclerotium against breast cancer cells in vitro |
| title_sort | molecular attributes and apoptosis inducing activities of a putative serine protease isolated from tiger milk mushroom lignosus rhinocerus sclerotium against breast cancer cells in vitro |
| topic | F5 Tiger milk mushroom Serine protease Lignosus rhinocerus Anticancer Apoptosis |
| url | https://peerj.com/articles/4940.pdf |
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