ISOLATION OF SULFATED POLYSACCHARIDES FROM GREEN ALGAE ENTEROMORPHA INTESTINALIS AND ITS EFFECT ON THE GROWTH OF HEPATOCARCINOMA CELL LINEAbstract
Identification of natural marine bioactive molecules and their therapeutic potential is valuable considering that there are reduced chances of undesirable side effects. This study was designed to isolate and purify the polysaccharides from E. intestinalis and evaluate its anti-cancer effect on human hepatocarcinoma cell lines (Hep3B). Polysaccharides from Enteromorpha intestinalis were isolated and purified. Preliminary characterization was done using UV-Vis spectrum and FTIR analysis. Total sulfate content of the polysaccharides was determined. MTT assay, AO / EB and DAPI staining were done to determine the cell growth pattern and nuclear changes effected by polysaccharides on Hep3B cells. Effect of polysaccharides on mitochondrial membrane potential and ROS production on Hep3B cells were evaluated using Rhodamine 123 and DCHF-DA staining. Results showed that polysaccharides from E. intestinalis were rich in sulfate groups (sPS). Cell viability, assessed in-vitro by MTT assay showed that the number of viable cells was less than 50% when exposed to concentrations exceeding 1 mg/ml of sPS. Cell viability and nuclear integrity assays depicted cells with compromised membrane integrity, cell damage, and nuclear perforation compared to control cells. Cells treated with sPS while stained with rhodamine 123 displayed a diffused pattern of staining with loss of intensity at higher concentrations indicating that loss of mitochondrial potential is a notable characteristic in sPS induced cytotoxicity. Hep-3B cells treated with sPS and stained with a ROS-specific fluorescent dye-DCFH-DA displayed cells with higher levels of ROS in comparison to control cells. These results may provide a basis on which further research in the development of marine drugs for the effective treatment of hepatocellular carcinoma.
A. G. Chandrika, R. Sivan, S. M. Chacko, P. K. Bhaskara and N. K. Govindan *
School of Biosciences, Mahatma Gandhi University, Kottayam, Kerala, India.
21 July 2017
25 August 2017
17 September 2017
01 January 2018