Please use this identifier to cite or link to this item:https://hdl.handle.net/20.500.12259/56799
Type of publication: Konferencijų tezės nerecenzuojamuose leidiniuose / Conference theses in non-peer-reviewed publications (T2)
Field of Science: Biologija / Biology (N010)
Author(s): Ayyappan, Priyadharishini;Ruzgys, Paulius;Mickevičius, Saulius;Šatkauskas, Saulius
Title: In vitro analysis of DNA damage of Bleomycin electrotransfered cells using Comet assay
Is part of: Smart Bio: international conference, 18-20 May 2017, Kaunas : abstracts book. Kaunas : Vytautas Magnus University, 2017
Extent: p. 92-92
Date: 2017
Keywords: Electroporation;DNA damage;Bleomycin;Comet assay;Cancer
ISBN: 9786098104424
Abstract: Currently, there is a need for individual, local tumor treatment with chemotherapeutical drugs. One of the methodologies for local anticancer drug delivery is electroporation. This phenomenon is induced when electric field is applied to the cells, by increasing transmembrane potential of the cell membrane. Temporal hydrophilic electropores are created in the membrane when electroporation threshold is reached. Created electropores are like bridge to hydrophilic anticancer drugs to enter affected cells. This way local (only electroporated cells) drug delivery method is obtained. At the moment local anticancer drug delivery via electroporation is available in clinics and is termed as electrochemotherapy. Mainly during electrochemotherapy treatment anticancer drug bleomycin (BLM) is used. This drug induces DNA breaks, causing cell death. However, to our knowledge there is no studies published, that indicate the quantification of DNA damage induced by BLM electrotransfer. Here we performed the in vitro analysis of DNA damage of BLM electrotransfered cells using the technique of Comet assay. Chinese hamster ovary (CHO) cell line was used as an object for BLM electrotransfer. Electroporation was performed with 1 HV (1400 V/cm) pulse. Used BLM concentration was from 2 mg/ml to 20 μg/ml. Electroporation was performed in electroporation medium (pH 7.1, conductivity 0.1 S/m, osmolarity 270 mOsm). Colonogenic assay was performed to evaluate cell viability. Electroporated cells were resuspended in low melting agarose (0.5 %), put on objective glass and covered with cover slip, 70 min after electroporation. Afterwards cells were kept in lysis buffer for 24 hours. Thereafter electrophoresis for 30 min in alkaline buffer (pH 13) was performed with voltage at 0.74 V/cm and 300 mA current. [...]
Internet: https://hdl.handle.net/20.500.12259/56799
Affiliation(s): Biologijos katedra
Fizikos katedra
Gamtos mokslų fakultetas
Vytauto Didžiojo universitetas
Appears in Collections:Universiteto mokslo publikacijos / University Research Publications

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