PT  - JOURNAL ARTICLE
AU  - EWA DROZD
AU  - JOLANTA KRZYSZTOŃ-RUSSJAN
AU  - BEATA GRUBER
TI  - Doxorubicin Treatment of Cancer Cells Impairs Reverse Transcription and Affects the Interpretation of RT-qPCR Results
DP  - 2016 Mar 01
TA  - Cancer Genomics - Proteomics
PG  - 161--170
VI  - 13
IP  - 2
4099  - http://cgp.iiarjournals.org/content/13/2/161.short
4100  - http://cgp.iiarjournals.org/content/13/2/161.full
SO  - Cancer Genomics Proteomics2016 Mar 01; 13
AB  - Background: Doxorubicin (DOX) acts in a variety of ways including DNA damage and enzyme inhibition, which consequently causes changes in gene expression of cells treated with this agent. Practical validation of experimental results followed by appropriate normalization of the factors investigated is crucial for obtaining biologically relevant results in gene expression studies. Materials and Methods: Six candidates were evaluated regarding their validity as internal reference genes: RPS23, FLOT2, UBB, ABCF1, ACTB, HPRT1. Optimization for quantitative polymerase reaction (qPCR) included: sensitivity, specificity, amplification efficiency and linear dynamic range determination. The gene expression stability was evaluated by real-time quantitative polymerase reaction (RT-qPCR) in two human cervical cancer cell lines: HeLa and DOX-resistant KB-V1 Cells treated under various concentrations of DOX. Results: DOX treatment changed gene expression and led to re-optimization of the cDNA template amounts. ACTB, HPRT1, RPS23 and FLOT2 are proposed to be sufficient as internal reference genes. Conclusion: DOX may alter the reverse transcription and amplification reactions of RT-qPCR, thus creating a risk of misinterpretation of gene expression results.