Apoptotic and Antiproliferative Potential of GAPDH from Mallotus philippensis Seed on Human Lung Carcinoma: In Vitro and In Vivo Approach

Aims: The anticancer potential of a purified seed protein from Mallotus philippensis is scientifically evaluated and reported here. Background: Seeds of Mallotus philippensis are used to treat various diseases in the indigenous systems of medicine in India. Objectives: The present study deals with the isolation, purification, identification, and screening of protein of interest that exhibit maximum activity against lung cancer cells from the seed crude protein of Mallotus philippensis. Methods: Size-exclusion with HPLC was used to purify crude protein (15 mg) from M. philippensis seeds. Protein of interest was identified using the LC-MS/MS method and analyzed by in vitro (A549 cell lines) in vivo (B16-F10 cells from melanoma cancer-induced Wistar rats) to estimate anticancer activity. Results: SDS-PAGE was applied to isolate and purify elution III (480 mu g/ml). Elution III LC-MS/MS data were used to search the UniProt database and were eventually matched with glyceraldehyde 3-phosphate dehydrogenase (GAPDH). MTT assay of GAPDH-treated A549 cells exhibited an IC50 of 3.03 +/- 0.39 mu g (24 h) and 1.93 +/- 0.19 mu g (48 h). AO/EtBr staining showed early and late apoptotic characteristics such as cell membrane blebbing, chromatin condensation, and the formation of apoptotic bodies. Hoechst staining confirmed the death of cells by exhibiting bright blue fluorescent, condensed, and fragmented nuclei. GAPDH-treated rats by 10 and 20 mg/kg bw significantly increased body weight by 29.50 +/- 3.06 and 31.33 +/- 2.69, respectively, and decreased melanoma metastasis in the lungs by 66.79% and 86.57%, respectively. Further, GAPDH treatment significantly increased the levels of SOD, CAT, and GPx and reduced GST and GSH. Histopathological analysis confirmed nuclear alteration in the lung tissue of the treated groups only. Conclusion: Apoptotic potential of GAPDH against lung carcinoma has been confirmed in the present investigation.