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@INPROCEEDINGS{Dahmen:255692,
author = {Dahmen, Volker and Kriehuber, Ralf},
title = {{I}nduction of the chromosomal translocation t(14;18) by
targeting the{BCL}-2 locus with specific binding
{I}-125-labeled {T}riplex-{F}orming oligonucleotides},
reportid = {FZJ-2015-05824},
year = {2015},
abstract = {Triplex-Forming oligonucleotides (TFO) are able to bind DNA
in a sequence specific manner and are a promising tool to
manipulate genes or gene regulatory units in a cellular
environment. TFO possess a therapeutic potential e.g. as a
carrier molecule for Alpha- or Auger-Electron-Emitter (AEE)
to target specific DNA sequences in tumor cells. We
established a method for the effective labeling of TFO with
the AEE Iodine-125 (I-125) and analyzed the influence of
I-125-labeled TFO in SCL-II cells on gene expression and
translocation frequency of the human BCL2 gene. The TFO-BCL2
employed in the present study binds to the BCL2 gene at
approximately 5400 bp upstream of the 3´-end. TFO labeling
with I-125 was performed using the primer extension method.
SCL-II cells were transfected with TFO via electroporation
and subsequently stored at -150°C for decay accumulation up
to a range from 100 to 330 decays/cell. SCL-II cells either
transfected with I-125-labeled multi-binding TFO (>300,000
targets per genome) or transfected with non-labeled TFO-BCL2
served as controls. Monitoring of BCL2 translocations was
done with the Fluorescence-In-Situ-Hybridization (FISH)
method. The utilized FISH probes were designed to detect a
translocation of the BCL2 gene from chromosome 18 to
chromosome 14, a common translocation found in follicular
lymphomas leading to an overexpression of BCL-2 protein.
Analysis of BCL2 gene expression levels was done via
quantitative Real-Time PCR on the Real Time PCR System 7500
(Applied Biosystems).The relative gene expression of BCl-2
in I-125-TFO-BCL2 transfected cells showed a significant
up-regulation when compared to the controls. Control SCL-II
cells were either transfected with I-125-labeled
multi-binding TFO (>300,000 targets per genome) or
non-labeled TFO-BCL2. Analysis of the BCL2 t(14;18)
translocation frequency revealed a significant 1.8- to
2-fold increase when compared to the control cells.We
conclude that I-125 decays within the BCL2 gene facilitates
the occurrence of the t(14;18) chromosomal translocation in
SCL-II cells and that the increased translocation frequency
of t(14;18) contributes to the observed overall enhanced
BCL-2 expression. However, it seems unlikely that these rare
translocation events fully explain the observed enhanced
gene expression of BCL-2.Funded by Bundesministerium für
Bildung und Forschung (BMBF), Grant 02NUK005A},
month = {May},
date = {2015-05-25},
organization = {15th International Congress of
Radiation Reasearch, Kyoto (Japan), 25
May 2015 - 29 May 2015},
subtyp = {After Call},
cin = {S-US},
cid = {I:(DE-Juel1)S-US-20090406},
pnm = {899 - ohne Topic (POF3-899)},
pid = {G:(DE-HGF)POF3-899},
typ = {PUB:(DE-HGF)24},
url = {https://juser.fz-juelich.de/record/255692},
}