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 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.

The Potential Role of Nrf2-PD-L1 Axis in Promoting of Oxaliplatin Resistance in Colon Cancer Cells

“…Duong et al also reported that Nrf2 silencing through siRNA promoted the e cacy of 5-FU in different pancreatic cancer cells including PANC-1, AsPC-1, and COLO-357 [53]. It has been demonstrated that a recombinant NRF2-siRNA signi cantly downregulated the NRF2-dependent ATP-binding cassette (ABC) e ux transporters, and increased the sensitivity of MG63 and 143B cells to sorafenib, doxorubicin, and cisplatin drugs [54]. Nrf2 suppression through siRNA increased ROS overwhelmed tumor growth and consequently promoted cell death through induction of sensitivity to chemotherapy [55].…”

Bioengineered miR-27b-3p and miR-328-3p modulate drug metabolism and disposition via the regulation of target ADME gene expression

“…Cytotoxicity studies were performed as described previously 31 . In brief, cells were seeded at 5000 cells/well in 96-well plate and cultured overnight.…”

Bioengineered Noncoding RNAs Selectively Change Cellular miRNome Profiles for Cancer Therapy

“…The recent termination of Phase I clinical study on synthetic miR-34a mimic (MRX34) (Beg et al, 2017), owing to high incidence of adverse immune responses, again testifies the body's capability to distinguish chemoengineered RNAi agents as foreign. Therefore, biologic approaches such as in vitro transcription (Beckert and Masquida, 2011) and, especially, bioengineering in live cells (Ponchon and Dardel, 2007;Ponchon et al, 2009;Huang et al, 2013;Li et al, 2014Li et al, , 2015Li et al, , 2018Chen et al, 2015;Wang et al, 2015;Pereira et al, 2016;Fang et al, 2017), are highly warranted to produce natural RNA molecules that should better represent cellular ncRNA properties for basic research and experimental therapy (Ho and Yu, 2016;Pereira et al, 2017).…”