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Published on: October 21, 2012
Gene expression analysis of cell death induction by taurolidine in different malignant cell lines
Ansgar M Chromik1, Stephan A Hahn, Adrien Daigeler
1Department of Visceral and General Surgery, St. Josef Hospital, Ruhr-University Bochum, Gudrunstrasse 56, D-44791 Bochum, Germany. a.chromik@klinikum-bochum.de
Background:
The anti-infective agent Taurolidine (TRD) has been shown to have cell death inducing properties, but the mechanism of its action is largely unknown. The aim of this study was to identify potential common target genes modulated at the transcriptional level following TRD treatment in tumour cell lines originating from different cancer types.
Methods:
Five different malignant cell lines (HT29, Chang Liver, HT1080, AsPC-1 and BxPC-3) were incubated with TRD (100 μM, 250 μM and 1000 μM). Proliferation after 8 h and cell viability after 24 h were analyzed by BrdU assay and FACS analysis, respectively. Gene expression analyses were carried out using the Agilent-microarray platform to identify genes which displayed conjoint regulation following the addition of TRD in all cell lines. Candidate genes were subjected to Ingenuity Pathways Analysis and selected genes were validated by qRT-PCR and Western Blot.
Results:
TRD 250 μM caused a significant inhibition of proliferation as well as apoptotic cell death in all cell lines. Among cell death associated genes with the strongest regulation in gene expression, we identified pro-apoptotic transcription factors (EGR1, ATF3) as well as genes involved in the ER stress response (PPP1R15A), in ubiquitination (TRAF6) and mitochondrial apoptotic pathways (PMAIP1).
Conclusions:
This is the first conjoint analysis of potential target genes of TRD which was performed simultaneously in different malignant cell lines. The results indicate that TRD might be involved in different signal transduction pathways leading to apoptosis.
Insights
Taurolidine (TRD) induces cancer cell death by regulating genes involved in apoptosis and ER stress. This study identified common target genes modulated by TRD across diverse tumor cell lines, revealing its potential in cancer therapy.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Taurolidine (TRD) exhibits cell death-inducing properties, but its precise mechanism of action remains largely unelucidated.
- Understanding TRD's molecular targets is crucial for its therapeutic development in oncology.
Purpose of the Study:
- To identify common target genes regulated at the transcriptional level by Taurolidine (TRD) treatment.
- To investigate TRD's mechanism of action in diverse cancer cell lines.
Main Methods:
- Incubation of five distinct malignant cell lines (HT29, Chang Liver, HT1080, AsPC-1, BxPC-3) with varying concentrations of TRD.
- Analysis of cell proliferation and viability using BrdU assay and FACS analysis.
- Gene expression profiling via microarray, followed by Ingenuity Pathways Analysis and validation using qRT-PCR and Western Blot.
Main Results:
- Taurolidine (TRD) at 250 μM significantly inhibited proliferation and induced apoptotic cell death in all tested cell lines.
- Key regulated genes included pro-apoptotic transcription factors (EGR1, ATF3), ER stress response genes (PPP1R15A), ubiquitination factors (TRAF6), and mitochondrial apoptosis pathway genes (PMAIP1).
Conclusions:
- This study presents the first conjoint analysis of Taurolidine's (TRD) potential target genes across multiple cancer cell types.
- The findings suggest that TRD modulates various signal transduction pathways, ultimately leading to apoptosis, highlighting its potential as an anti-cancer agent.
