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

Yap, Hui Yeng Yeannie and Tan, Nget Hong and Ng, Szu Ting and Tan, Chon Seng and Fung, Shin Yee (2018) 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, 6. e4940. ISSN 2167-8359, DOI https://doi.org/10.7717/peerj.4940.

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Official URL: https://doi.org/10.7717/peerj.4940

Abstract

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 nonsynonymous 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.

Item Type: Article
Funders: Ministry of Education Malaysia: Fundamental Research Grant Scheme (FRGS) FP029-2014A, MAHSA University: MAHSA Research Grant (RP122-09/17), University of Malaya, Kuala Lumpur, Malaysia: UMRG Programme Grant (RP034 A, B, C–17AFR)
Uncontrolled Keywords: Anticancer; Apoptosis; F5; GME4347_g; Lignosus rhinocerus; Serine protease; Tiger milk mushroom
Subjects: R Medicine
Divisions: Faculty of Medicine
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 14 Feb 2019 07:40
Last Modified: 14 Feb 2019 07:40
URI: http://eprints.um.edu.my/id/eprint/20316

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