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Cellular Oncology

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01-12-2013 | Original Paper

siRNA-mediated knock-down of DFF45 amplifies doxorubicin therapeutic effects in breast cancer cells

Authors: Fatemeh Bagheri, Shahrokh Safarian, Mohamadreza Baghaban Eslaminejad, Nader Sheibani

Published in: Cellular Oncology | Issue 6/2013

Abstract

Purpose

RNA interference (RNAi) has become a promising tool for cancer therapy. Small interfering RNAs (siRNAs) can synergistically enhance the cell killing effects of drugs used in cancer treatment. Here we examined the effects of siRNA-mediated DNA fragmentation factor 45 (DFF45) gene silencing on breast cancer cell viability, cell cycle arrest, and apoptosis in the presence and absence of doxorubicin.

Methods

We designed three siRNAs, which target different regions of the DFF45 mRNA. Gene silencing was confirmed by real time RT-PCR and Western blot analyses. The impact of DFF45 siRNA, doxorubicin, and their combination on the viability, cell cycle and apoptosis of T-47D and MDA-MB-231 breast cancer cells were determined by MTT, PI staining, annexin V binding, caspase-3 activity, DNA laddering, and chromatin condensation assays.

Results

Based on flow cytometric analyses, we found that silencing of DFF45 alone had little effect on apoptosis, especially in T-47D cells. However, when used in combination with doxorubicin (0.33 μM) a significant increase (P < 0.05) in apoptosis was observed in T-47D and MDA-MB-231 cells, i.e., ~2.5- and 3-fold, respectively. Caspase-3 activity, chromatin condensation, as well as DNA laddering supported increased apoptosis in the combinatorial treatment. Cell cycle arrest in both cell lines occurred at lower levels after siRNA + doxorubicin treatment compared to doxorubicin only.

Conclusions

Our data indicate that DFF45 gene silencing, when applied in combination with doxorubicin, may offer a novel therapeutic strategy for the treatment of breast cancer.

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Title: siRNA-mediated knock-down of DFF45 amplifies doxorubicin therapeutic effects in breast cancer cells
Authors: Fatemeh Bagheri
Shahrokh Safarian
Mohamadreza Baghaban Eslaminejad
Nader Sheibani
Publication date: 01-12-2013
Publisher: Springer Netherlands
Published in: Cellular Oncology / Issue 6/2013
Print ISSN: 2211-3428
Electronic ISSN: 2211-3436
DOI: https://doi.org/10.1007/s13402-013-0157-1

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