Bioengineered NRF2-siRNA is effective to interfere with NRF2 pathways and improve chemosensitivity of human cancer cells

Peng Cheng Li, Mei Juan Tu, Pui Yan Ho, Joseph L. Jilek, Zhijian Duan, Qian Yu Zhang, Ai Xi Yu, Aiming Yu

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The nuclear factor (erythroid-derived 2)-like 2 (NRF2) is a transcription factor in the regulation of many oxidative enzymes and efflux transporters critical for oxidative stress and cellular defense against xenobiotics. NRF2 is dysregulated in patient osteosarcoma (OS) tissues and correlates with therapeutic outcomes. Nevertheless, research on the NRF2 regulatory pathways and its potential as a therapeutic target is limited to the use of synthetic small interfering RNA (siRNA) carrying extensive artificial modifications. Herein, we report successful high-level expression of recombinant siRNA against NRF2 in Escherichia coli using our newly established noncoding RNA bioengineering technology, which was purified to >99% homogeneity using an anion-exchange fast protein liquid chromatography method. Bioengineered NRF2-siRNA was able to significantly knock down NRF2 mRNA and protein levels in human OS 143B and MG63 cells, and subsequently suppressed the expression of NRF2-regulated oxidative enzymes [heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1] and elevated intracellular levels of reactive oxygen species. In addition, recombinant NRF2-siRNA was effective to sensitize both 143B and MG63 cells to doxorubicin, cisplatin, and sorafenib, which was associated with significant downregulation of NRF2-targeted ATP-binding cassette (ABC) efflux transporters (ABCC3, ABCC4, and ABCG2). These findings support that targeting NRF2 signaling pathways may improve the sensitivity of cancer cells to chemotherapy, and bioengineered siRNA molecules should be added to current tools for related research.

Original languageEnglish (US)
Pages (from-to)2-10
Number of pages9
JournalDrug Metabolism and Disposition
Volume46
Issue number1
DOIs
StatePublished - Jan 1 2018

Fingerprint

Small Interfering RNA
Osteosarcoma
Neoplasms
Antiporters
Bioengineering
Untranslated RNA
Heme Oxygenase-1
ATP-Binding Cassette Transporters
Xenobiotics
Enzymes
Research
Liquid Chromatography
NAD
Doxorubicin
Cisplatin
Reactive Oxygen Species
Oxidoreductases
Oxidative Stress
Transcription Factors
Down-Regulation

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

Cite this

Bioengineered NRF2-siRNA is effective to interfere with NRF2 pathways and improve chemosensitivity of human cancer cells. / Li, Peng Cheng; Tu, Mei Juan; Ho, Pui Yan; Jilek, Joseph L.; Duan, Zhijian; Zhang, Qian Yu; Yu, Ai Xi; Yu, Aiming.

In: Drug Metabolism and Disposition, Vol. 46, No. 1, 01.01.2018, p. 2-10.

Research output: Contribution to journalArticle

Li, Peng Cheng ; Tu, Mei Juan ; Ho, Pui Yan ; Jilek, Joseph L. ; Duan, Zhijian ; Zhang, Qian Yu ; Yu, Ai Xi ; Yu, Aiming. / Bioengineered NRF2-siRNA is effective to interfere with NRF2 pathways and improve chemosensitivity of human cancer cells. In: Drug Metabolism and Disposition. 2018 ; Vol. 46, No. 1. pp. 2-10.
@article{6e0ac69482ed4fbc8115f36553a38d4c,
title = "Bioengineered NRF2-siRNA is effective to interfere with NRF2 pathways and improve chemosensitivity of human cancer cells",
abstract = "The nuclear factor (erythroid-derived 2)-like 2 (NRF2) is a transcription factor in the regulation of many oxidative enzymes and efflux transporters critical for oxidative stress and cellular defense against xenobiotics. NRF2 is dysregulated in patient osteosarcoma (OS) tissues and correlates with therapeutic outcomes. Nevertheless, research on the NRF2 regulatory pathways and its potential as a therapeutic target is limited to the use of synthetic small interfering RNA (siRNA) carrying extensive artificial modifications. Herein, we report successful high-level expression of recombinant siRNA against NRF2 in Escherichia coli using our newly established noncoding RNA bioengineering technology, which was purified to >99{\%} homogeneity using an anion-exchange fast protein liquid chromatography method. Bioengineered NRF2-siRNA was able to significantly knock down NRF2 mRNA and protein levels in human OS 143B and MG63 cells, and subsequently suppressed the expression of NRF2-regulated oxidative enzymes [heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1] and elevated intracellular levels of reactive oxygen species. In addition, recombinant NRF2-siRNA was effective to sensitize both 143B and MG63 cells to doxorubicin, cisplatin, and sorafenib, which was associated with significant downregulation of NRF2-targeted ATP-binding cassette (ABC) efflux transporters (ABCC3, ABCC4, and ABCG2). These findings support that targeting NRF2 signaling pathways may improve the sensitivity of cancer cells to chemotherapy, and bioengineered siRNA molecules should be added to current tools for related research.",
author = "Li, {Peng Cheng} and Tu, {Mei Juan} and Ho, {Pui Yan} and Jilek, {Joseph L.} and Zhijian Duan and Zhang, {Qian Yu} and Yu, {Ai Xi} and Aiming Yu",
year = "2018",
month = "1",
day = "1",
doi = "10.1124/dmd.117.078741",
language = "English (US)",
volume = "46",
pages = "2--10",
journal = "Drug Metabolism and Disposition",
issn = "0090-9556",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "1",

}

TY - JOUR

T1 - Bioengineered NRF2-siRNA is effective to interfere with NRF2 pathways and improve chemosensitivity of human cancer cells

AU - Li, Peng Cheng

AU - Tu, Mei Juan

AU - Ho, Pui Yan

AU - Jilek, Joseph L.

AU - Duan, Zhijian

AU - Zhang, Qian Yu

AU - Yu, Ai Xi

AU - Yu, Aiming

PY - 2018/1/1

Y1 - 2018/1/1

N2 - The nuclear factor (erythroid-derived 2)-like 2 (NRF2) is a transcription factor in the regulation of many oxidative enzymes and efflux transporters critical for oxidative stress and cellular defense against xenobiotics. NRF2 is dysregulated in patient osteosarcoma (OS) tissues and correlates with therapeutic outcomes. Nevertheless, research on the NRF2 regulatory pathways and its potential as a therapeutic target is limited to the use of synthetic small interfering RNA (siRNA) carrying extensive artificial modifications. Herein, we report successful high-level expression of recombinant siRNA against NRF2 in Escherichia coli using our newly established noncoding RNA bioengineering technology, which was purified to >99% homogeneity using an anion-exchange fast protein liquid chromatography method. Bioengineered NRF2-siRNA was able to significantly knock down NRF2 mRNA and protein levels in human OS 143B and MG63 cells, and subsequently suppressed the expression of NRF2-regulated oxidative enzymes [heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1] and elevated intracellular levels of reactive oxygen species. In addition, recombinant NRF2-siRNA was effective to sensitize both 143B and MG63 cells to doxorubicin, cisplatin, and sorafenib, which was associated with significant downregulation of NRF2-targeted ATP-binding cassette (ABC) efflux transporters (ABCC3, ABCC4, and ABCG2). These findings support that targeting NRF2 signaling pathways may improve the sensitivity of cancer cells to chemotherapy, and bioengineered siRNA molecules should be added to current tools for related research.

AB - The nuclear factor (erythroid-derived 2)-like 2 (NRF2) is a transcription factor in the regulation of many oxidative enzymes and efflux transporters critical for oxidative stress and cellular defense against xenobiotics. NRF2 is dysregulated in patient osteosarcoma (OS) tissues and correlates with therapeutic outcomes. Nevertheless, research on the NRF2 regulatory pathways and its potential as a therapeutic target is limited to the use of synthetic small interfering RNA (siRNA) carrying extensive artificial modifications. Herein, we report successful high-level expression of recombinant siRNA against NRF2 in Escherichia coli using our newly established noncoding RNA bioengineering technology, which was purified to >99% homogeneity using an anion-exchange fast protein liquid chromatography method. Bioengineered NRF2-siRNA was able to significantly knock down NRF2 mRNA and protein levels in human OS 143B and MG63 cells, and subsequently suppressed the expression of NRF2-regulated oxidative enzymes [heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1] and elevated intracellular levels of reactive oxygen species. In addition, recombinant NRF2-siRNA was effective to sensitize both 143B and MG63 cells to doxorubicin, cisplatin, and sorafenib, which was associated with significant downregulation of NRF2-targeted ATP-binding cassette (ABC) efflux transporters (ABCC3, ABCC4, and ABCG2). These findings support that targeting NRF2 signaling pathways may improve the sensitivity of cancer cells to chemotherapy, and bioengineered siRNA molecules should be added to current tools for related research.

UR - http://www.scopus.com/inward/record.url?scp=85038410785&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85038410785&partnerID=8YFLogxK

U2 - 10.1124/dmd.117.078741

DO - 10.1124/dmd.117.078741

M3 - Article

C2 - 29061583

AN - SCOPUS:85038410785

VL - 46

SP - 2

EP - 10

JO - Drug Metabolism and Disposition

JF - Drug Metabolism and Disposition

SN - 0090-9556

IS - 1

ER -