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Applied Biochemistry and Biotechnology

, Volume 187, Issue 4, pp 1539–1550 | Cite as

Investigation of Vipera Anatolica Venom Disintegrin via Intracellular Uptake with Radiolabeling Study and Cell-Based Electrochemical Biosensing Assay

  • Ozge Er
  • Ece Eksin
  • Hale Melis Soylu
  • Bayram Göçmen
  • Ayşe NalbantsoyEmail author
  • Fatma YurtEmail author
  • Arzum ErdemEmail author
Article

Abstract

Snake venoms are a natural biological source that has potential therapeutic value with various protein compounds. Disintegrins originally were discovered as a family of proteins from snake venoms composed of cysteine rich low molecular weight polypeptides. Disintegrins exhibit specific binding and higher affinity toward integrin with potential inhibition of function. Trans-membrane receptors of the integrin family may involve in many pathological conditions such as inflammation and tumor progression with important processes related to invasion and migration. Since disintegrins have the ability to bind to integrins, they could be used for cancer detection and treatment, and in monitoring of therapy in select cancer types. The main purpose of the study is to investigate disintegrin containing Vipera anatolica (VAT) crude venom potential for radiolabeling and intracellular uptake as well as electrochemical biosensing assay against U87MG human brain glioblastoma cells. For this purpose, VAT crude venom containing U87MG cell-specific disintegrin was investigated in terms of radiolabeling and intracellular uptake as well as electrochemical biosensing assay in comparison with echistatin (ECT) disintegrin in cells. The interaction between VAT crude venom and ECT with HEK293 human non-tumorigenic embryonic kidney cells and glioblastoma U87MG cells was electrochemically investigated using pencil graphite electrodes (PGEs). The interaction of the VAT crude venom and ECT with HEK293 and U87MG cells was detected according to the changes in oxidation signals. Then, VAT crude venom and echistatin were labeled with 131I via iodogen method. Intracellular uptakes of radiolabeled molecules were investigated in U87MG cell line. 131I-VAT can be an agent for imaging of glioblastoma cancer. Further work will focus on the production of large quantities of pure VAT disintegrin with a biotechnological approach to improving imaging agent.

Keywords

Vipera anatolica crude venom Glioblastoma cancer Cancer imaging Electrochemical biosensing assay 

Abbreviations

VAT

Vipera anatolica

U87MG

human brain glioblastoma cells

ECT

echistatin

HEK293

human non-tumorigenic embryonic kidney cells

PGEs

pencil graphite electrodes

ECM

extracellular matrix

RDG

Arg-Gly-Asp

ROS

reactive oxygen species

VEGF

vascular endothelial growth factor

DPV

differential pulse voltammetry

TLRC

thin layer radio chromatography

BCA

bicinchoninic acid

DMEM/F12

Dulbecco’s modified Eagle’s medium F12

FBS

fetal bovine serum

MTT

3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide)

OD

optical density

IC50

half maximal inhibition of growth

PBS

phosphate buffer solution

RIPA

radio-immunoprecipitation assay

ITLC-cellulose

cellulose-coated plastic

Notes

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Nuclear Applications, Institute of Nuclear SciencesEge UniversityIzmirTurkey
  2. 2.Department of Analytical Chemistry, Faculty of PharmacyEge UniversityIzmirTurkey
  3. 3.The Institute of Natural and Applied Sciences, Biotechnology DepartmentEge UniversityIzmirTurkey
  4. 4.The Institute of Natural and Applied Sciences, Biomedical Technology DepartmentEge UniversityIzmirTurkey
  5. 5.Zoology Section, Department of Biology, Faculty of ScienceEge UniversityIzmirTurkey
  6. 6.Department of Bioengineering, Faculty of EngineeringEge UniversityIzmirTurkey

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