$Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species$

Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species

Soft corals are sessile marine invertebrates having vast number of promising anticancer compounds. Various studies have proven the effectiveness of soft coral extract on various cancer cell lines including liver cancer cell lines. Currently, highthroughput metabolomics is applied to replace conven...

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Main Author:	Lim, Leong Rui
Format:	Thesis
Language:	English
Published:	2018
Subjects:	SH Aquaculture Fisheries Angling
Online Access:	https://eprints.ums.edu.my/id/eprint/25765/1/Identification%20of%20liver%20cancer%20%28Hep%2038%29%20inhibiting%20fraction%20from%20soft%20coral%20Sinularia%20Species.pdf
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spelling	my-ums-ep.257652020-08-07T07:31:23Z Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species 2018 Lim, Leong Rui SH Aquaculture. Fisheries. Angling Soft corals are sessile marine invertebrates having vast number of promising anticancer compounds. Various studies have proven the effectiveness of soft coral extract on various cancer cell lines including liver cancer cell lines. Currently, highthroughput metabolomics is applied to replace conventional technique in drug discovery which is time-consuming and laborious. The current study aimed to determine the efficiency of a selected soft coral's fraction, SCJJ-FS against hepatocellular carcinoma Hep 3B and its cell death triggering activities using mass spectrometry based metabolomics. Sixteen soft corals were collected and extracted using modified Folch extraction protocol. Soft corals' crude extracts were screened against the hepatocellular carcinoma (Hep 3B) cells. The most effective crude extracts of SCOB and SCJJ were further isolated using chromatography technique, producing eight fractions for each crude extract. These fractions were acquired and tested at different concentrations (25, 50 and 100 µg/ml) for 48 h. SCJJ-FSwas found to be the most effective in inducing cell death in Hep 3B. The fraction was screened with MTT assay to evaluate cell proliferation. From the assay, the lowest inhibitory concentration at 50% of total cell survival (ICso) was determined at 45.31 µg/ml at 72 h post-treatment. Following, cell metabolomics was applied at ICso against SCJJ-FS and then underwent the extraction process. Toe cell crude extracts were subjected to high-throughput profiling using ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight. Chemometric analysis revealed over 50 metabolites were significantly (p-value < 0.05) perturbed and these metabolites were found involving in the programmed cell death via tandem mass spectral matching. Up-regulation of vitamin D and its derivatives increased the concentration of pro-apoptotic factor, calcium ions (Ca2•) which caused direct programmed cell death. For triacylglycerols, their concentration was high in the treated Hep 3B cells that might cause distortion in plasma membrane. For diaclyglycerols, they act as second lipid messenger to activate proteins in cellular activities. On the other hand, perturbation of phospholipids such as phosphatidylcholine and phosphatidylethanolamine affected the rigidity of plasma membrane and regulation of signaling molecules synthesis and cell death. Up-regulation of ceramides caused programmed cell death by disrupting mitochondria while down-regulated ceramides were involved in synthesis of sphingomyelins in which sphingomyelins helped to recover plasma membrane. To conclude, SCJJ-FS has cytotoxicity effect towards Hep 3B cell line. In the future, a structural identification of the SC11-F5 is needed. 2018 Thesis https://eprints.ums.edu.my/id/eprint/25765/ https://eprints.ums.edu.my/id/eprint/25765/1/Identification%20of%20liver%20cancer%20%28Hep%2038%29%20inhibiting%20fraction%20from%20soft%20coral%20Sinularia%20Species.pdf text en public mphil masters Universiti Malaysia Sabah Biotechnology Research Institute
institution	Universiti Malaysia Sabah
collection	UMS Institutional Repository
language	English
topic	SH Aquaculture Fisheries Angling
spellingShingle	SH Aquaculture Fisheries Angling Lim, Leong Rui Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species
description	Soft corals are sessile marine invertebrates having vast number of promising anticancer compounds. Various studies have proven the effectiveness of soft coral extract on various cancer cell lines including liver cancer cell lines. Currently, highthroughput metabolomics is applied to replace conventional technique in drug discovery which is time-consuming and laborious. The current study aimed to determine the efficiency of a selected soft coral's fraction, SCJJ-FS against hepatocellular carcinoma Hep 3B and its cell death triggering activities using mass spectrometry based metabolomics. Sixteen soft corals were collected and extracted using modified Folch extraction protocol. Soft corals' crude extracts were screened against the hepatocellular carcinoma (Hep 3B) cells. The most effective crude extracts of SCOB and SCJJ were further isolated using chromatography technique, producing eight fractions for each crude extract. These fractions were acquired and tested at different concentrations (25, 50 and 100 µg/ml) for 48 h. SCJJ-FSwas found to be the most effective in inducing cell death in Hep 3B. The fraction was screened with MTT assay to evaluate cell proliferation. From the assay, the lowest inhibitory concentration at 50% of total cell survival (ICso) was determined at 45.31 µg/ml at 72 h post-treatment. Following, cell metabolomics was applied at ICso against SCJJ-FS and then underwent the extraction process. Toe cell crude extracts were subjected to high-throughput profiling using ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight. Chemometric analysis revealed over 50 metabolites were significantly (p-value < 0.05) perturbed and these metabolites were found involving in the programmed cell death via tandem mass spectral matching. Up-regulation of vitamin D and its derivatives increased the concentration of pro-apoptotic factor, calcium ions (Ca2•) which caused direct programmed cell death. For triacylglycerols, their concentration was high in the treated Hep 3B cells that might cause distortion in plasma membrane. For diaclyglycerols, they act as second lipid messenger to activate proteins in cellular activities. On the other hand, perturbation of phospholipids such as phosphatidylcholine and phosphatidylethanolamine affected the rigidity of plasma membrane and regulation of signaling molecules synthesis and cell death. Up-regulation of ceramides caused programmed cell death by disrupting mitochondria while down-regulated ceramides were involved in synthesis of sphingomyelins in which sphingomyelins helped to recover plasma membrane. To conclude, SCJJ-FS has cytotoxicity effect towards Hep 3B cell line. In the future, a structural identification of the SC11-F5 is needed.
format	Thesis
qualification_name	Master of Philosophy (M.Phil.)
qualification_level	Master's degree
author	Lim, Leong Rui
author_facet	Lim, Leong Rui
author_sort	Lim, Leong Rui
title	Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species
title_short	Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species
title_full	Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species
title_fullStr	Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species
title_full_unstemmed	Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species
title_sort	identification of liver cancer (hep 38) inhibiting fraction from soft coral sinularia species
granting_institution	Universiti Malaysia Sabah
granting_department	Biotechnology Research Institute
publishDate	2018
url	https://eprints.ums.edu.my/id/eprint/25765/1/Identification%20of%20liver%20cancer%20%28Hep%2038%29%20inhibiting%20fraction%20from%20soft%20coral%20Sinularia%20Species.pdf
_version_	1747836555008933888

Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species

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