Effect Of Fucoidan From Undaria Pinnatifida On Human Lung Adenocarcinoma Cancer A549Cells: Anti-proliferation, Anti-migration And Induction Apoptosis

Objective: Fucoidan, known as fucose glycans or sulfate polysaccharides, it isalso a necessary substance of brown algae to use light and nutrients. Fucoidan is awater-soluble polysaccharide mixture including L-fucose and sulfate as maincomposition, galactose, xylose, glucose and glucuronic acid. Fucoidan has a variety ofbiological activity. Recent researches indicate that in vitro and vivo experiment,fucoidan presents a significant anti-proliferation activity on many kinds of tumor cells.It will contribute to the development of new drugs and functional products. This articleaims to explore the apoptosis pathway of human lung adenocarcinoma A549cellswhich induced by fucoidan extracted from Undaria pinnatifida.Method:1. Acquiring fucoidan sample by the hot water extraction and ethanolfractionation method from U.pinnatifida sporophyll powder. Sulfuric acid-phenolassay referred to the sugar content. Barium sulfate turbidimetric method referred to thesulfate content. Sulfuric acid-carbazole colorimetric assay referred to the uronic acids.2. Using MTT colorimetric to quantify cell vitality of A549cells after beingtreated with fucoidan, MTT assay of A549cells adhesion inhibition, Transwell chamberassay on A549cells migration inhibition.3. Immunizing animals for prepare the Fucoidan antibodies, to detect serumantibodies by ELISA and immunofluorescence.4. Determining expression of apoptosis related protein Caspase-3in A549cells byWestern Blot, spectrophotometer detection of Caspase-8, Caspase-9enzyme activitychanges; RT-PCR detection of Survivin mRNA expression.Results:1. This fucoidan sample consisted mainly of carbohydrates (59.2%)，sulfates (21%) and uronic acid (9.13%) with fucose and galactose constituting themonosaccharide composition. The optical rotation of the fucoidan (0.6mg/ml) showed a value of0.99°at20°C.2. The proliferation of A549cells was inhibited by fucoidan. The adhesion andmigration of A549cells are also significantly inhibited.3. ELISA results showed that the serum of tests group contain antibody forFucoidan. Immunofluorescence method validates the presence of antibodies for it.4. Results of Western Blot and enzyme activity test indicate that fucoidan canup-regulate the activity of Caspase-3,Caspase-8,Caspase-9in A549cells by time anddose-dependent manner. Results of RT-PCR show that Fucoidan can down-regulationof Survivin mRNA expression.Conclusions:1. Fucoidan can significantly inhibit human lung adenocarcinomacell line A549proliferation, adhesion and migration.2. Fucoidan induced apoptosis of A549cells through the Fas-associated signalingpathway and the mitochondrial pathway.