Apoptosis induced by nonspecific effects of siRNA in human umbilical vein endothelial cell

Hyean Woo Lee, Sun Ju Choi, Yeong Min Yoo, Joon Hyung Sohn, Sang Baek Koh, Kyu Sang Park

Research output: Contribution to journalArticle

Abstract

RNA interference (RNAi) is a powerful tool used to produce post-translational gene silencing in a sequence-dependent manner. However, sequence-independent activation of interferon systems induced by double-stranded RNA can interfere with the data interpretation of RNAi experiments. In this study, we observed cytokine activation and cytotoxicity caused by small interfering RNA (siRNA) in human umbilical vein endothelial cells (HUVECs). Transfection with different sequences of siRNAs (21-nucleotides) induced apoptosis in HUVECs within 24 hours, in a dose-dependent and sequence-independent manner. These effects could not be achieved by the application of transfection agents nor by siRNA only. In HUVECs, the expression of toll-like receptor 3 (TLR3), a receptor for double-stranded RNA, was significantly increased by the transfection with siRNAs regardless of their sequences. The expressions of interleukin 6, interleukin 8, and interferon-β were also up-regulated by siRNA transfection. The activation of RNA-dependent protein kinase (PKR) peaked within 30 min and slowly decreased 4 hours after transfection with siRNA, which was followed by the phosphorylation of eukaryotic initiation factor 2α(eIF2α). These studies suggest that transfection with double-stranded oligonucleotides in HUVECs shows sequence-independent apoptotic effects associated with the upregulation of TLR3 and cytokines, which should be considered when designing experiments using RNAi.

Original languageEnglish
Pages (from-to)50-56
Number of pages7
JournalToxicology and Environmental Health Sciences
Volume4
Issue number1
DOIs
Publication statusPublished - 2012 Dec 1

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Endothelial cells
Human Umbilical Vein Endothelial Cells
Small Interfering RNA
Transfection
Toll-Like Receptor 3
eIF-2 Kinase
Apoptosis
Double-Stranded RNA
Chemical activation
RNA
RNA Interference
Interferons
Eukaryotic Initiation Factor-2
Cytokines
Phosphorylation
Cytotoxicity
Interleukin-8
Oligonucleotides
Interleukin-6
Nucleotides

All Science Journal Classification (ASJC) codes

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

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title = "Apoptosis induced by nonspecific effects of siRNA in human umbilical vein endothelial cell",
abstract = "RNA interference (RNAi) is a powerful tool used to produce post-translational gene silencing in a sequence-dependent manner. However, sequence-independent activation of interferon systems induced by double-stranded RNA can interfere with the data interpretation of RNAi experiments. In this study, we observed cytokine activation and cytotoxicity caused by small interfering RNA (siRNA) in human umbilical vein endothelial cells (HUVECs). Transfection with different sequences of siRNAs (21-nucleotides) induced apoptosis in HUVECs within 24 hours, in a dose-dependent and sequence-independent manner. These effects could not be achieved by the application of transfection agents nor by siRNA only. In HUVECs, the expression of toll-like receptor 3 (TLR3), a receptor for double-stranded RNA, was significantly increased by the transfection with siRNAs regardless of their sequences. The expressions of interleukin 6, interleukin 8, and interferon-β were also up-regulated by siRNA transfection. The activation of RNA-dependent protein kinase (PKR) peaked within 30 min and slowly decreased 4 hours after transfection with siRNA, which was followed by the phosphorylation of eukaryotic initiation factor 2α(eIF2α). These studies suggest that transfection with double-stranded oligonucleotides in HUVECs shows sequence-independent apoptotic effects associated with the upregulation of TLR3 and cytokines, which should be considered when designing experiments using RNAi.",
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Apoptosis induced by nonspecific effects of siRNA in human umbilical vein endothelial cell. / Lee, Hyean Woo; Choi, Sun Ju; Yoo, Yeong Min; Sohn, Joon Hyung; Koh, Sang Baek; Park, Kyu Sang.

In: Toxicology and Environmental Health Sciences, Vol. 4, No. 1, 01.12.2012, p. 50-56.

Research output: Contribution to journalArticle

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AU - Koh, Sang Baek

AU - Park, Kyu Sang

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