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

Lim, Leong Rui (2018) Identification of liver cancer (Hep 38) inhibiting fraction from soft coral Sinularia species. Masters thesis, Universiti Malaysia Sabah.

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Abstract

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.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Soft corals , marine , metabolomics
Subjects: S Agriculture > SH Aquaculture. Fisheries. Angling
Divisions: SCHOOL > Biotechnology Research Institute (BRI)
Depositing User: Noraini
Date Deposited: 07 Aug 2020 07:31
Last Modified: 07 Aug 2020 07:31
URI: http://eprints.ums.edu.my/id/eprint/25765

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