title: Anticancer activity of a low immunogenic protein toxin (BMP1) from Indian toad (Bufo melanostictus, Schneider) skin extract.
abstract: Earlier, a protein (BMP1, MW-79kDa) had been isolated from Indian toad (Bufo melanostictus) skin aqueous extract possessed anticancer activity against EAC bearing mice (Bhattacharjee et al., 2011). In the present study, the anti-proliferative and apoptogenic activities of BMP1 have been evaluated in leukemic (U937 and K562) and hepatoma (HepG2) cells. BMP1 dose dependently inhibited U937 and K562 cell growth having IC₅₀ values of 49 μg/ml and 30 μg/ml respectively. The anti-proliferative activity of BMP1 was observed in MTT assay, proliferating cell nuclear antigen (PCNA) expression and cell cycle arrest study. Flow-cytometric data revealed that BMP1 arrested cell cycle in U937 and K562 cells at Sub-G1 and G1 phases. The BMP1-induced dose dependent expressions of CDKIs (p21(cip1) and p27(kip1)) and inhibition of CDK2 and PCNA expression in HepG2 cells support the inhibition of cell proliferation due to G1 arrest. BMP1-induced apoptosis analyzed by annexin-V binding study and the DNA fragmentation by comet assay were correlated with the sub-G1 arrest. The parallel induction of bax and p53 expression in HepG2 cells and the up-regulation of caspase 3 and caspase 9 due to BMP1 treatment indicated the involvement of p53-dependent intrinsic pathway of apoptosis. BMP1 was found to be low immunogenic in nature.
authors.affiliation: Laboratory of Toxinology and Experimental Pharmacodynamics, University of Calcutta, Department of Physiology, 92 Acharya Prafulla Chandra Road, Kolkata 700009, India. gomesantony@hotmail.com
authors.forename: Antony
authors.initials: A
authors.lastname: Gomes
authors.forename: Biplab
authors.initials: B
authors.lastname: Giri
authors.forename: Aftab
authors.initials: A
authors.lastname: Alam
authors.forename: Sanghamitra
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Pushpak
authors.initials: P
authors.lastname: Bhattacharjee
authors.forename: Aparna
authors.initials: A
authors.lastname: Gomes
mesh_terms: D029845:Amphibian Proteins / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002025:Bufonidae; D053148:Caspase 3 / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D020552:Comet Assay; D054697:Cytostatic Agents / Q000302:isolation & purification / Q000494:pharmacology*; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D008297:Male; D051379:Mice; D011817:Rabbits; D014020:Tissue Extracts / Q000737:chemistry*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D029845:Amphibian Proteins; D017304:Annexin A5; D000970:Antineoplastic Agents; C494826:BAX protein, human; D018797:Cell Cycle Proteins; D054697:Cytostatic Agents; D014020:Tissue Extracts; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D053148:Caspase 3