In-vitro cytotoxicity and apoptosis mechanism of ethyl acetate extract from Trichoderma reesei strain TV221 associated with marine sponge: Stylissa flabelliformis

Erna Prawita Setyowati; Sylvia Utami Tunjung Pratiwi; Purwantiningsih; Indah Purwantini

doi:10.7324/JAPS.2018.8921

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Abstract

This research aimed to acknowledge the cytotoxic effect of ethyl acetate extract fermented from Trichoderma reesei strain TV221 fungi associated with Stylissa flabelliformis sponge in various cell lines such as WiDr, T41, T47D, Raji, and Vero, as well as to discover the mechanism of cell’s death. The result of fermented fungi T. reesei was extracted with ethyl acetate. Ethyl acetate extract was tested against cell lines using the MTT assay. The result of cytotoxic selectivity test with lowest IC50 price, then apoptosis test with double staining acridine orange-ethidium bromide method was performed Ethyl acetate extract fermented from T. reesei has cytotoxic ability against tumor cell lines (WiDr, T41, T47D, Raji) and is not toxic to normal cell lines (Vero). The test sample can spur apoptosis process on colon cancer cells Widr.

Keyword: Trichoderma reesei strain TV221 cytotoxic effect apoptosis Stylissa flabelliformis associated sponge.

Citation:

Setyowati EP, Pratiwi SUT, Purwantiningsih, Purwantini I. Invitro cytotoxicity and apoptosis mechanism of ethyl acetate extract from Trichoderma reesei strain TV221 associated with marine sponge: Stylissa flabelliformis. J App Pharm Sci, 2018; 8(09): 151-157.

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Badisa RB, Reed SFD, Josph P, Cooperhood JS, Latinwo LM, Goodman CB. Selective Cytotoxic Activities of Two Novel Synthetic Drugs on Human Breast Carcinoma MCF-7 Cells. Anticancer Res, 2009; 29(8):2993-2996.
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Proksch P, Edrada RA, Ebel R. Drugs from The Seas–Current Status and Microbiological Implications. Applied Microbiology and Biotechnology, 2002; 125-134.
Setyowati EP, Sudarsono dan Wahyuono S, Cytotoxicity and antimicrobial test of the bioactive compound isolated from Stylissa flabelliformis sponge. Indonesian Journal of Pharmacy, 2004, 15(2):50-56.
Setyowati EP, Sylvia UTP, Purwantiningsih, Samara O. Bioactivity of Fungi Trichoderma reesei Associated with Sponges Stylissa flabelliformis Collected from National Park West Bali, Indonesia. Journal of Biological Sciences, 2017; 17(8):362-368. https://doi.org/10.3923/jbs.2017.362.368
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Setyowati1 E P [PubMed] [Google Scholar ]

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Pratiwi1 S U T [PubMed] [Google Scholar ]

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Almeida AP, Dethoup T, Singburaudon N, Lima R, Vasconcelos MH, Pinto M, Kijjoa A. The in vitro anticancer activity of the crude extract of the sponge-associated fungus Eurotium cristatumand its secondary metabolites. Journal of Natural Pharmaceuticals, 2010; 1(1).

Badisa RB, Reed SFD, Josph P, Cooperhood JS, Latinwo LM, Goodman CB. Selective Cytotoxic Activities of Two Novel Synthetic Drugs on Human Breast Carcinoma MCF-7 Cells. Anticancer Res, 2009; 29(8):2993-2996.

Brinkman CM, Marker A, Kurtböke DI. An Overview on Marine Sponge-Symbiotic Bacteria as Unexhausted Sources for Natural Product Discovery, Diversity, 2017; 9(40).

CCRC. Cytotoxic test protocol with MTT assay, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta. 2013.

CLSI, Methods for Dilution Antimicrobial Susceptibility Test for Bacteria that Grow Aerobically; Approved Standard, 7th Edition, Clinical and Laboratory Standards Institute, Wayne, Pensylvania. 2006.

Drzewiecka D. Significance and Roles of Proteus spp. Bacteria in Natural Environments. Microb Ecol, 2016; 72:741-758. https://doi.org/10.1007/s00248-015-0720-6

Elmore S. Apoptosis: A Review of Programmed Cell Death. Toxicol Pathol, 2007; 35(4):495-516. https://doi.org/10.1080/01926230701320337

Fink SL, Cookson BT. Apoptosis, Pyroptosis, and Necrosis: Mechanistic Description of Dead and Dying Eukaryotic Cells. Infect Immun, 2005; 73(4):1907-1916. https://doi.org/10.1128/IAI.73.4.1907-1916.2005

Hooper JNA, Sponguide: Guide to Sponge Collection and Identification, Queensland Museum, South Brisbane. 2004.

Kennedy J, Marchesi JR, Dobson ADW. Metagenomic Approaches to Exploit the Biotechnological Potential of the Microbial Consortia of Marine Sponges. Applied Microbiology and Biotechnology, 2007; 75(1):11-20. https://doi.org/10.1007/s00253-007-0875-2

Levrero M, De Laurenzi V, Constanzo A, Sabatini S, Gong J, Wang JYJ, Melino G. The p53/p63/p73 Family of Transcription Factors: Overlapping and Distinct Functions. Journal of Cell Science, 2000; 113:1661-1670.

Liu HC, Chen GG, Vlantis AC, Leung BCS, Tong MCE, van Hasselt CA. 5-Fluorouracil Mediates Apoptosis and G1/S Arrest in Laryngeal Squamous Cell Carcinoma via a p53-Independent Pathway. The Cancer Journal, 2006; 12(6):482-49. https://doi.org/10.1097/00130404-200611000-00008

Norbury CJ, Hickson ID, Cellular responses to DNA damage. Annu Rev Pharmacol Toxicol, 2001; 41:367-401. https://doi.org/10.1146/annurev.pharmtox.41.1.367

Proksch P, Edrada RA, Ebel R. Drugs from The Seas–Current Status and Microbiological Implications. Applied Microbiology and Biotechnology, 2002; 125-134.

Setyowati EP, Sudarsono dan Wahyuono S, Cytotoxicity and antimicrobial test of the bioactive compound isolated from Stylissa flabelliformis sponge. Indonesian Journal of Pharmacy, 2004, 15(2):50-56.

Setyowati EP, Sylvia UTP, Purwantiningsih, Samara O. Bioactivity of Fungi Trichoderma reesei Associated with Sponges Stylissa flabelliformis Collected from National Park West Bali, Indonesia. Journal of Biological Sciences, 2017; 17(8):362-368. https://doi.org/10.3923/jbs.2017.362.368

Setyowati EP, Sylvia UTP, Purwantiningsih, Samirana PO. Antimicrobial activity and Identification of fungus associated Stylissa flabelliformis sponge collected from Menjangan Island West Bali National Park. Indonesia, Indonesian Journal of Pharmacy, 2018; 29(2):66-73. https://doi.org/10.14499/indonesianjpharm29iss2pp66

Sutejo IR, Putri H, Meiyanto E. The Selectivity of Ethanolic Extract of Buah Makassar (Brucea javanica) on In vitro Study of Metastatic Breast Cancer. Journal of Agromedicine and Medical Sciences, 2016; 2(1):1-5. https://doi.org/10.19184/ams.v2i1.2422

Taylor MW, Radax R, Steger D, Wagner M. Sponge-Associated Microorganisms: Evolution, Ecology, and Biotechnological Potential. Microbiol Mol Biol Rev, 2007; 71(2):295–347. https://doi.org/10.1128/MMBR.00040-06

Thomas TRA, Kavlekar DP, Lokabharathi PA. Marine Drugs from Sponge–Microbe Association – A Review. Marine Drugs, 2010, 8(4): 1417–146. https://doi.org/10.3390/md8041417

Thoms C, Schupp PJ. Chemical defense strategies in sponges: a review, Porifera research: Biodiversity. Innovation and sustainability, 2006; 627-637.

Wang G. Diversity and biotechnological potential of the sponge-associated microbial consortia. Journal of Industrial Microbiology and Biotechnology, 2006; 33(7):545-551. https://doi.org/10.1007/s10295-006-0123-2

Weerapreeyakul N, Nonpunya A, Barusrux S, Thitimetharoch T, Sripanidkulchai B. Evaluation of the Anticancer Potential of the Six Herbs Againts a Hepatoma Cell Line. Chinese Medicine Journal, 2012; 7:15. https://doi.org/10.1186/1749-8546-7-15

Dogan SM, Ercetin AP, Altun Z, Dursun D, Aktas S. Gene expression characteristic of breast cancer stem cell. JBUON, 2015; 20:1304-1313.

Ebad SS, Proksch P. Bioactive Secondary Metabolites from Marine Derived Fungi in Marine Pharmacognosy Trends and Applications edited by Se Kwon Kim. CRC, 2013; 27-52.