A heat stable protein toxin (drCT-I) from the Indian Viper (Daboia russelli russelli) venom having antiproliferative, cytotoxic and apoptotic activities.
A heat stable 7.2kDa protein toxin (drCT-I) has been purified and crystallized from Indian Daboia russelli russelli venom (Roy Choudhury et al., 2006. Acta Cryst. F Struct Biol Cryst Commun, 62(Pt. 3), 292). The N-terminal (first 20) amino acid sequence of drCT-I was LKCNKLVPLFYKTCPAGKNL, which showed sequence homology to cytotoxins isolated from Naja venom. drCT-I has been evaluated for anticancer activity against EAC cells in vivo and human leukemic cells (U937, K562) in vitro. drCT-I (125 microg/kg, i.p/day for 10 days) significantly decreased EAC cell count, cell viability (p<0.001) and significantly increased the survival time of tumour bearing mice (T/C% 178.64, p<0.01) in comparison to untreated tumour bearing control. drCT-I, produced dose and time-dependent inhibition of U937 and K562 cell growth and had an IC50 of 8.9 and 6.7 microg/ml respectively after 24h treatment. The reduced MTT values after drCT-I treatment indicated its cytotoxic nature, which supported its antiproliferative action. Scanning electron microscopy and confocal microscopy in U937 and K562 cells after drCT-I treatment indicated certain features of apoptosis such as membrane blebbing, perforations, nuclear fragmentation. The induction of apoptosis was further confirmed by phosphatidylserine externalization observed using annexinV-FITC/PI staining and flow cytometric analysis. drCT-I brought about apoptosis by G1 phase arrest of the cell cycle. The effect of drCT-I on normal human peripheral blood mononuclear cell (PBMNC) viability and cytotoxicity was studied in culture and was found to be lower than that on U937 and K562 cells. Thus both in vivo and in vitro experimental results suggested that drCT-I possessed anticancer potential.
Gomes
Antony
A
Laboratory of Toxinology and Experimental Pharmacodynamics, Department of Physiology, University of Calcutta, 92 A. P. C. Road, Kolkata 700 009, India. agomes_cu@rediffmail.com
Choudhury
Subhasree Roy
SR
Saha
Archita
A
Mishra
R
R
Giri
Biplab
B
Biswas
A K
AK
Debnath
Anindita
A
Gomes
Aparna
A
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor / Q000187:drug effects / Q000648:ultrastructure; D049109:Cell Proliferation / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D020128:Inhibitory Concentration 50; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D014757:Viper Venoms / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D017819:Viperidae
D000970:Antineoplastic Agents; D014757:Viper Venoms