Alteration in mitochondrial superoxide production by shRNA gene silencing of AIF in various cell line

VAŘECHA, Miroslav, Daniela PÁCLOVÁ, Jiřina PROCHÁZKOVÁ, Pavel MATULA and Michal KOZUBEK. Alteration in mitochondrial superoxide production by shRNA gene silencing of AIF in various cell line. In Advances in Molecular and Cancer Bilogy. 2010. ISBN 978-80-210-5312-0.

Basic information

• Original name: Alteration in mitochondrial superoxide production by shRNA gene silencing of AIF in various cell line
• Name in Czech: Změny v produkci mitochondriálního kyslíkového radikálu způsobené genovým utišením AIF pomocí shRNA v různých buněčných liniích
• Authors: VAŘECHA, Miroslav (203 Czech Republic, guarantor, belonging to the institution), Daniela PÁCLOVÁ (203 Czech Republic), Jiřina PROCHÁZKOVÁ (203 Czech Republic, belonging to the institution), Pavel MATULA (203 Czech Republic, belonging to the institution) and Michal KOZUBEK (203 Czech Republic, belonging to the institution).
• Edition: Advances in Molecular and Cancer Bilogy, 2010.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: Genetics and molecular biology
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• RIV identification code: RIV/00216224:14330/10:00045338
• Organization unit: Faculty of Informatics
• ISBN: 978-80-210-5312-0
• Keywords in English: AIF; shRNA; gene silencing; image analysis; superoxide; mitochondria; oxidative phosphorylation
• Tags: International impact

Abstract

Apart from this function in cell death, AIF also plays largely unknown role in cell life. We used AIF gene silencing using shRNA vectors followed by measurements by flow cytometry and confocal fluorescent microscopy to detect the superoxide production during oxidative phosphorylation in mitochondria. We managed to knock-down the expression of gene AIF by transient or stable transfection in various human cell lines. Interestingly, mitochondrial superoxide production in various cell lines reacted to gene silencing of AIF differently. Cell line U-2 OS transiently and also stably transfected with vector shRNA was found to produce less superoxide in comparison to control U-2 OS cells. Interestingly, HeLa cell line shows opposite results with significantly increased mitochondrial superoxide production in silenced HeLa cells. We are also in process of examination of other healthy and cancer cell. Conclusions about possible role of AIF in mitochondria based on our results with various AIF-silenced cell lines will be presented.

Links

• LC535, research and development project: Name: Dynamika a organizace chromosomů během buněčného cyklu v normě a patologii

Investor: Ministry of Education, Youth and Sports of the CR, Dynamika a organizace chromosomů během buněčného cyklu v normě a patologii

• MSM0021622419, plan (intention): Name: Vysoce paralelní a distribuované výpočetní systémy

Investor: Ministry of Education, Youth and Sports of the CR, Highly Parallel and Distributed Computing Systems

• 2B06052, research and development project: Name: Vytipování markerů, screening a časná diagnostika nádorových onemocnění pomocí vysoce automatizovaného zpracování multidimenzionálních biomedicínských obrazů (Acronym: Biomarker)

Investor: Ministry of Education, Youth and Sports of the CR, Determination of markers, screening and early diagnostics of cancer diseases using highly automated processing of multidimensional biomedical images