EXPLORING DOUBLE‐STRANDED RNA AS A POSSIBLE CHEMOTHERAPEUTIC AGENT
Danielle N. Schramm, Sandrine Lépine, Jessica K. Bell
Abstract
Danielle N. Schramm, Sandrine Lépine, Jessica K. Bell
Abstract
Pattern recognition receptors (PRRs) function in the innate immune system to alert the host to pathogenic components. Double‐stranded RNA (dsRNA), viral genomic material or a viral replication intermediate, trigger the PRRs toll‐like receptor 3 (TLR3), retinoic acid‐inducible gene I (RIG‐I), melanoma differentiation‐associated gene 5 (MDA5), and dsRNA‐dependent protein kinase receptor (PKR). Previous studies have reported that dsRNA stimulation induced TLR3‐dependent apoptosis. Objectives Examine 786‐O, OVCAR‐3, OVCA‐420, SKOV‐3, DOV‐13, CAOV‐3 cells for dsRNA receptor expression and determine dsRNA‐induced response. Methods PRR expression was confirmed by RT‐PCR. Expression levels were determined via qPCR. Apoptotic response to dsRNA stimulation was measured by Hoechst staining 48h post‐stimulation. Results 786‐O, SKOV‐3 and DOV‐13 show no response to dsRNA. RT‐PCR analysis showed that cell lines contained mRNA for all dsRNA receptors. qPCR showed a significant increase in PRR expression in those cell lines that underwent dsRNA‐induced apoptosis. Conclusions Four dsRNA receptors are present in our cell lines. dsRNA receptor expression does not correlate with dsRNA induced apoptosis, but increased dsRNA receptor expression suggests receptor levels or altered signaling pathways may reduce sensitivity to dsRNA. Funding provided by National Cancer Institute & Jimmy V Foundation.
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Pattern recognition receptors (PRRs) function in the innate immune system to alert the host to pathogenic components. Double‐stranded RNA (dsRNA), viral genomic material or a viral replication intermediate, trigger the PRRs toll‐like receptor 3 (TLR3), retinoic acid‐inducible gene I (RIG‐I), melanoma differentiation‐associated gene 5 (MDA5), and dsRNA‐dependent protein kinase receptor (PKR). Previous studies have reported that dsRNA stimulation induced TLR3‐dependent apoptosis. Objectives Examine 786‐O, OVCAR‐3, OVCA‐420, SKOV‐3, DOV‐13, CAOV‐3 cells for dsRNA receptor expression and determine dsRNA‐induced response. Methods PRR expression was confirmed by RT‐PCR. Expression levels were determined via qPCR. Apoptotic response to dsRNA stimulation was measured by Hoechst staining 48h post‐stimulation. Results 786‐O, SKOV‐3 and DOV‐13 show no response to dsRNA. RT‐PCR analysis showed that cell lines contained mRNA for all dsRNA receptors. qPCR showed a significant increase in PRR expression in those cell lines that underwent dsRNA‐induced apoptosis. Conclusions Four dsRNA receptors are present in our cell lines. dsRNA receptor expression does not correlate with dsRNA induced apoptosis, but increased dsRNA receptor expression suggests receptor levels or altered signaling pathways may reduce sensitivity to dsRNA. Funding provided by National Cancer Institute & Jimmy V Foundation.
Key concepts: RNA silencing, MDA5, TLR3, Biology, Receptor, RIG-I, Protein kinase R, Molecular biology