Small interfering RNA-mediated suppression of serum response factor, E2-promotor binding factor and survivin in non-small cell lung cancer cell lines by non-viral transfection.

Abstract

OBJECTIVES:

Serum response factor (SRF), E2F1 and survivin are well-known factors involved in a multitude of cancer-related regulation processes. However, to date, no suitable means has been found to apply their potential in the therapy of non-small cell lung cancer (NSCLC). This study deals with questions of small interfering ribonucleic acid (siRNA) transfection efficiency by a non-viral transfection of NSCLC cell-lines and the power of siRNA to transiently influence cell division by specific silencing.

METHODS:

Different NSCLC cell lines were cultured under standard conditions and transfected, with specific siRNA targeting SRF, E2F1 and survivin in a non-viral manner. Cells treated with non-specific siRNA (SCR-siRNA) served as controls. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed for messenger RNA (mRNA) expression levels. Additionally, transfection efficiency was evaluated by flow cytometry. The analysis of cell proliferation was determined with a CASY cell counter 3 days after transfection with SRF or SCR-siRNA.

RESULTS:

Transfection of the NSCLC cell lines with specific siRNAs against SRF, E2F1 and survivin resulted in a very considerable reduction of the intracellular mRNA concentration. CASY confirmation of cell viability demonstrated an excellent survival of the cell lines treated with non-specific siRNA, in contrast to with application of specific siRNA.

CONCLUSIONS:

This study reports a reliable transfectability of NSCLC-cell lines by siRNA, initially in a non-viral manner, and a reproducible knockdown of the focussed targets, consequently leading to the death of the tumour cells. This constitutes a strong candidate for a new assessment strategy in the therapy of non-small cell lung cancer.

• Authors: Tobias Walker, Andrea Nolte, Volker Steger, Christina Makowiecki, Migdat Mustafi, Godehard Friedel, Christian Schlensak, Hans-Peter Wendel
• Journal: European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery (Eur J Cardiothorac Surg) Vol. 43 Issue 3 Pg. 628-33; discussion 633-4 (Mar 2013) ISSN: 1873-734X [Electronic] Germany
• PMID: 23152437 (Publication Type: Journal Article)

Chemical References

• BIRC5 protein, human
• E2F1 Transcription Factor
• E2F1 protein, human
• Inhibitor of Apoptosis Proteins
• RNA, Messenger
• RNA, Small Interfering
• SRF protein, human
• Serum Response Factor
• Survivin

Topics

• Carcinoma, Non-Small-Cell Lung (genetics, metabolism, therapy)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• E2F1 Transcription Factor (genetics, metabolism)
• Gene Silencing
• Genetic Therapy (methods)
• Humans
• Inhibitor of Apoptosis Proteins (genetics, metabolism)
• Lung Neoplasms (genetics, metabolism, therapy)
• RNA, Messenger (genetics, metabolism)
• RNA, Small Interfering (administration & dosage, genetics)
• Serum Response Factor (genetics, metabolism)
• Survivin
• Transfection