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@INPROCEEDINGS{Dahmen:865160,
author = {Dahmen, Volker and Schmitz, Sabine and Kriehuber, Ralf},
title = {{I}nduction of specific chromosomal rearrangements by
targeting sensitive genomic loci using{I}-125-labeled
{T}riplex-forming oligonucleotides},
reportid = {FZJ-2019-04706},
year = {2019},
abstract = {Triplex-Forming-oligonucleotides (TFO) bind DNA in a
sequence-specific manner and possess therapeutic potential
e.g. as a carrier-molecule for Auger-Electron-Emitter (AEE)
targeting specific DNA sequences in tumor cells. We
established a method for the labeling of TFO with the AEE
Iodine-125 (125I) and analyzed the influence of 125I-labeled
TFO in SCL-II cells on gene expression, translocation
frequency and protein expression of the human BCL2 gene.
TFO-BCL2 binds to the BCL2 gene upstream of the 3´-end. TFO
labeling with 125I 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. SCL-II cells transfected with
125I-labeled multi-binding TFO or non-labeled TFO-BCL2
served as controls. Monitoring of BCL2 translocations was
done with Fluorescence-In-Situ-Hybridization (FISH). The
utilized FISH-probes were designed to detect a
characteristic t(14;18) translocation of the BCL2 gene,
leading to an overexpression of BCL-2. Gene expression
levels were measured via qRT-PCR. Verification of gene
expression on the protein level was analyzed by Western
blotting. The relative gene expression of BCl-2 in
125I-TFO-BCL2 transfected cells showed a significant
up-regulation and analysis of the BCL2 t(14;18)
translocation frequency revealed a significant ~ 1.8-fold
increase. This increase was not reflected on the protein
level.We conclude that 125I decays within the BCL2 gene
facilitate the t(14;18) chromosomal translocation and that
the increased translocation frequency contributes to the
observed enhanced BCL-2 gene expression. Funded by
Bundesministerium für Bildung und Forschung (BMBF), Grant
02NUK005A and 02NUK043A},
month = {Sep},
date = {2019-09-09},
organization = {51st Annual Meeting of the Association
for Radiation Protection, Würzburg
(Germany), 9 Sep 2019 - 12 Sep 2019},
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/865160},
}