Aim:
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 isolation, purification, identification and screening protein of interest that exhibits 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 LC-MS/MS method and analyzed by in-vitro (A549 cell lines) in-vivo (B16-F10 cells melanoma cancer-induced Wistar rats) to estimate anticancer activity.
Results:
SDS-PAGE was applied to isolate and purify elution III (480 μg/ml). LC-MS/MS data of the elution III were used for searching UniProt database and finally matched with glyceraldehyde 3-phosphate dehydrogenase (GAPDH). MTT assay of GAPDH-treated A549 cells exhibited IC50 of 3.03 ± 0.39 µg (24 h) and 1.93±0.19 µg (48 h). AO/EtBr staining showed early and late apoptotic characters such as cell membrane blebbing, chromatin condensation and 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 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 is confirmed in the present investigation.
