Anti-proliferation and apoptotic effect of Scylla serrata extract soup on human acute lymphoblastic leukemia cell line (MOLT-4)

Chemical compounds derived from natural products have been used to treat human diseases from the dawn of ancient medicine. Mangrove crab, Scylla serrata is a euryhaline species from Portunidae family that inhabits intertidal zones and estuaries of Africa, Australia and Asia. It possess chitin, chitosan and selenium which have shown nontoxic and contain functional properties in lowering the rates of blood cancer. The aim of the study was to evaluate in-vitro anti-proliferative and apoptotic effect of S. serrata extract soup targeting on human acute lymphoblastic leukemia cell line (MOLT-4). Apoptosis has been considered as a promising key target for cancer chemoprevention and there are numerous experiments involve apoptosis-inducing agents which play important role in either initiation or execution of apoptosis. Cytotoxicity of the S. serrata extract soup on MOLT-4 cell was assayed using standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric procedure. The anti-proliferative and apoptotic effects of S. serrata extract soup in MOLT-4 cells were tested via reverse transcriptase polymerase chain reaction (RT-PCR), quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) using selective apoptotic-related gene marker Fas, Caspase-3, p38-MAPKs, p53 and housekeeping gene β-actin. Detection of apoptosis and distinguish cell population were assessed via flow cytometry assay. The result obtained from the cytotoxicity assay showed the significant reduction of cytotoxic effect after 72 hours of treatment at the dose of more than 25% (v/v) with inhibitory concentration of 50% cell population (IC50) at concentration 35% (v/v) in comparison with the normal cells. The effect is in a dose and time-dependent manner. Anti-proliferative and apoptotic effect of S. serrata extract soup on MOLT-4 cells using selective gene markers are showed to increase expression of pro-apoptotic gene markers Fas, p38-MAPKs and p53 except for Caspase-3 gene suggesting that apoptosis process is not through pathway of Caspase-3. Based on Annexin V-FITC/Propidium Iodide flow cytometric assay, population of viable MOLT-4 cells reduced up to 35% and undergo both early and late apoptosis when treated with S. serrata extract soup 35% (v/v) at 72 hours incubation. This suggests that S. serrata extract soup has an in-vitro anti-proliferative and apoptotic effect on human acute lymphoblastic leukemia cell line (MOLT-4) and could be potential use as an alternative therapy or supplementary diet for leukemia.