Abstract
Scorpion and its organs have been used to cure epilepsy, rheumatism, and male impotency since medieval times. Scorpion venom which contains different compounds like enzyme and non-enzyme proteins, ions, free amino acids, and other organic inorganic substances have been reported to posses antiproliferative, cytotoxic, apoptogenic, and immunosuppressive properties. We for the first time report the apoptotic and antiproliferative effects of scorpion venom (Odontobuthus doriae) in human neuroblastoma cells. After exposure of cells to medium containing varying concentrations of venom (10, 25, 50, 100, and 200 μg/ml), cell viability decreased to 90.75, 75.53, 55.52, 37.85, and 14.30%, respectively, after 24 h. Cells expressed morphological changes like swelling, inhibition of neurite outgrowth, irregular shape, aggregation, rupture of membrane, and release of cytosolic contents after treatment with venom. Lactate dehydrogenase (LDH) level increased in 50 and 100 μg/ml as compared to control, but there was no significant increase in LDH level at a dose of 10 and 20 μg/ml. Two concentrations viz. 50 and 100 μg/ml were selected because of the profound effect of these concentrations on the cellular health and population. Treatment with these two concentrations induced reactive nitrogen intermediates and depolarization in mitochondria. While caspase-3 activity increased in a concentration-dependent manner, only 50 μg/ml was able to fragment DNA. It was interesting to note that at higher dose, i.e., 100 μg/ml, the cells were killed, supposedly by acute necrosis. DNA synthesis evidenced by bromodeoxyuridine (BrdU) incorporation was inhibited in a concentration-dependent manner. The cells without treatment incorporated BrdU with high affinity confirming their cancerous nature whereas very less incorporation was noticed in treated cells. Our results show apoptotic and antiproliferative potential of scorpion venom (O. doriae) in human neuroblastoma cells. These properties make scorpion venom a valuable therapeutic agent in cancer research.
Similar content being viewed by others
References
Costa-N EM (2005) Animal based medicines: biological prospection and the sustainable of zootherapeutic resources. Ann Brazil Acad Sci 77:33–34
Wang WX, Ji YH (2005) Scorpion venom induces glioma cell apoptosis in vivo and inhibits glioma tumor growth in vitro. J Neurooncol 73:1–7
Kuhn-Nentwig L (2003) Antimicrobial and cytolytic peptides of venomous arthropods. CMLS 60:2651–2668
Incesu Z, Calıskan F, Zeytinoglu H (2005) Cytotoxic and gelatinolytic activities of Mesobuthus Gibbosus (Brullé, 1832) venom. Revista CENIC Ciencias Biológicas 36
Possani LD, Becerril B, Delepierre M, Tytgat J (1999) Scorpion toxins specific for Na+_ channels. Eur J Biochem 264:287–300
DeBin JA, Maggio JE, Strichartz GR (1993) Purification and characterization of chlorotoxin, a chloride channel ligand from the venom of the scorpion. Am J Physiol 264:361–369
Gallagher JD, Fay MJ, North WG, McCann FV (1996) Ionic signals in T47D human breast cancer cells. Cell Signal 8:279–284
Laniado ME, Fraser SP, Djamgoz MB (2001) Voltage gated K(+) channel activity in human prostrate cancer cell lines of markedly different metastatic potential: distinguishing characteristics of PC-3 and LNCaP cells. Prostate 46:262–274
Das Gupta S, Debnath A, Saha A, Giri B, Tripathi G, Vedasiromoni JR, Gomes A, Gomes A (2007) Indian black scorpion (Heterometrus bengalensis Koch) venom induced antiproliferative and apoptogenic activity against human leukemic cell lines U937 and K562. Leukemia Res 31:817–825
Omran MAA (2003) Cytotoxic and apoptotic effects of scorpion Leiurus quinquestriatus venom on 293T and C2C12 eukaryotic cell line. J Venom Anim Toxin incl Trop Dis 9:255–276
Omran MAA (2003) In vitro Anticancer effect of scorpion Leiurus quinquestriatus and Egyptian Cobra venom on human breast and prostate cancer cell lines. J Med Sci 3:66–68
Soroceanu L, Gillespie Y, Khazaeli MB, Sontheimer H (1998) Use of chlorotoxin for targeting of primary brain tumors. Cancer Res 58:4871–4879
Ismail M, El-Asmar MF, Osman OH (1975) Pharmacological studies with scorpion (Palamneus gravimanus) venom: evidence for the presence of histamine. Toxicon 13:49–50
Shiau Lin SY, Tseng WC, Lee CY (1975) Pharmacology of scorpion toxin II in the skeletal muscle. J Biomed Life Sci 289:359–368
Matos IM, Teixeira MM, Leite R, Freire-Maia L (1999) Pharmacological evidence that neuropeptides mediate part of the action of scorpion venom on the guinea pig ileum. Eur J Pharmacol 368:231–236
Schwartz EF, Capes EM, Diego-García E, Zamudio Fernando Z, Fuentes O, Possani Lourival D, Valdivia Hector H (2009) Characterization of hadrucalcin, a peptide from Hadrurus gertschi scorpion venom with pharmacological activity on ryanodine receptors. Br J Pharmacol 157:392–403
Adolfo B, Sylvia S, Huub JOdC, Elena V, Marco A, Marcelo JMA, Alicia J, Leonardo DS, Olinda D (2006) In vitro leishmanicidal activity of Tityus discrepans scorpion venom. Parasitol Res 99:167–173
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63
Decker T, Lohmann-M ML (1988) A quick and simple method for the quantitation of lactate hydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity. J Immunol Methods 15:61–69
Ding AH, Nathan CF, Stuehr DJ (1988) Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages: comparison of activating cytokines and evidence for independent production. J Immunol 141:2407–2412
Smiley ST, Reers M, Mottola-H C, Lin M, Chen A, Smith TW, Steele GD, Chen LB (1991) Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate forming lipophilic cation JC-1. Proc Natl Acad Sci USA 88:3671–3675
Kweon SM, Lee ZW, Yi SJ, Kim YM, Han JA, Paik SG, Ha KS (2004) Protective role of tissue transglutaminase in the cell death induced by TNF-α in SH-SY5Y neuroblastoma cells. J Biochem Mol Biol 37:185–191
Dover R, Patel K (1994) Improved methodology for detecting bromodeoxyuridine in cultured cells and tissue sections by immunocytochemistry. Histochemistry 102:383–387
Gadwalkar SR, Bushan S, Pramod K, Gouda C, Kumar PM (2006) Bilateral cerebellar infarction: a rare complication of scorpion sting. JAPI 54:581–583
Chow G, Kini RM (2001) Exogenous factors from animal sources that induce platelet aggregation. Thromb Haemost 85:177–178
Sugahara T, Takahashi T, Yamaya S, Ohsaka A (1976) In vitro aggregation of platelets induced by alpha-toxin (phospholipase C) of Clostridium perfringens. Jpn J Med Sci Biol 29:255–263
Zhou L, Zhu DY (2009) Neuronal nitric oxide synthase: structure, subcellular localization, regulation, and clinical implications. Nitric Oxide 20:223–230
Finocchietto PV, Franco MC, Holod S, Gonzalez AS, Converso DP, Arciuch VG, Serra MP, Poderoso JJ, Carreras MC (2009) Mitochondrial nitric oxide synthase: a masterpiece of metabolic adaptation, cell growth, transformation, and death. Exp Biol Med 234:1020–1028
Leonelli M, Torrao AS, Britto LR (2009) Unconventional neurotransmitters, neurodegeneration and neuroprotection. Braz J Med Biol Res 42:68–75
Grune T, Reinheckel T, North JA, Ruili Bescos PB, Shringarpure R, Davies KJA (2002) Ezrin turnover and cell shape changes catalyzed by proteasome in oxidatively stressed cells. FASEB J 16:1602–1610
Vieira H, Kroemer G (2003) Mitochondria as targets of apoptosis regulation by nitric oxide. IUBMB Life 55:613–616
Caroppi P, Sinibaldi F, Fiorucci L, Santucci R (2009) Apoptosis and human diseases: mitochondrion damage and lethal role of released cytochrome C as proapoptotic protein. Curr Med Chem 16:4058–4065
Kroemer G, Galluzzi L, Brenner C (2007) Mitochondrial membrane permeabilization in cell death. Physiol Rev 87:99–163
Cole K, Perez-Polo JR (2004) Neuronal trauma model: in search of Thanatos. Int J Dev Neurosci 22:485–496
Zhang YY, Wu LC, Wang ZP, Wang ZX, Jia Q, Jiang GS, Zhang WD (2009) Anti-proliferation effect of polypeptide extracted from scorpion venom on human prostate cancer cells in vitro. J Clin Med Res 1:24–31
Acknowledgments
Authors are thankful to Dr. G.N Qazi (Vice Chancellor, Jamia Hamdard) for his continued support during this study. Authors also express their gratitude to NCCS, Pune for timely shipment of the cell lines. Department of Biochemistry acknowledges the support of DST to establish infrastructure for animal tissue culture facility under FIST program.
Conflict of interest
The authors declare that there are no conflicts of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zargan, J., Sajad, M., Umar, S. et al. Scorpion (Odontobuthus doriae) venom induces apoptosis and inhibits DNA synthesis in human neuroblastoma cells. Mol Cell Biochem 348, 173–181 (2011). https://doi.org/10.1007/s11010-010-0652-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-010-0652-x