Apoptosis-Inducing Active Protein from Marine Clam Donax variabilis on NSCLC Cells

Abstract

The marine environment has been represented as the most vital source of bioactive constituents. In the present work, we intended to purification, characterization, and investigation of the apoptotic effect of the bioactive molecule from edible mollusc Donax variabilis on NSCLC cell lines. The bioactive molecules were purified using Anion Exchange Chromatographic method with different millimolar of NaCl concentrations. The fractions were further performed to check the proliferative effect on A549 and NCI-H23 cells by MTT assay. The molecular mass of the active purified protein fraction (PPF-V) was 40 kDa, determined by SDS-PAGE. Furthermore, MALDI-TOF/MS was disclosed that it shared a 91% protein sequence similarity with FMRFamide peptide. Meanwhile, the apoptotic effect of PPF-V on A549 and NCI-H23 cells were investigated. Immunoblot analysis found that the PPF-V treatment groups showed the upregulation of Bax, Cytochrome-c, Cleaved Caspase-9 and 3 expression and downregulation of Bcl-2 protein expression on both NSCLC cell lines. In addition, purified protein fraction induced the loss of mitochondrial membrane potential and generation of reactive oxygen species (ROS) on lung cancer cells. Altogether, our results proved that the PPF-V from Donax variabilis inducing apoptosis against NSCLC cell lines, which could be a potential natural candidate for lung cancer therapy.