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

被引:7
作者
Walker, Tobias [1 ]
Nolte, Andrea [1 ]
Steger, Volker [1 ]
Makowiecki, Christina [1 ]
Mustafi, Migdat [1 ]
Friedel, Godehard [2 ]
Schlensak, Christian [1 ]
Wendel, Hans-Peter [1 ]
机构
[1] Univ Tubingen Hosp, Dept Thorac Cardiac & Vasc Surg, Tubingen, Germany
[2] Schillerhohe Hosp, Dept Thorac Surg, Stuttgart, Germany
关键词
Lung cancer; siRNA; SRF; E2F1; Survivin; ADHESION MOLECULES; ENDOTHELIAL-CELLS; APOPTOSIS; PROTECTION; PATHWAY; GENE;
D O I
10.1093/ejcts/ezs337
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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. 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. 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. 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.
引用
收藏
页码:628 / 634
页数:7
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