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Repression of caspase-3 and RNA-binding protein HuR cleavage by cyclooxygenase-2 promotes drug resistance in oral squamous cell carcinoma

Oncogene volume 36, pages 3137–3148 (2017)Cite this article

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Abstract

A well-studied RNA-binding protein Hu Antigen-R (HuR), controls post-transcriptional gene regulation and undergoes stress-activated caspase-3 dependent cleavage in cancer cells. The cleavage products of HuR are known to promote cell death; however, the underlying molecular mechanisms facilitating caspase-3 activation and HuR cleavage remains unknown. Here, we show that HuR cleavage associated with active caspase-3 in oral cancer cells treated with ionizing radiation and chemotherapeutic drug, paclitaxel. We determined that oral cancer cells overexpressing cyclooxygenase-2 (COX-2) limited the cleavage of caspase-3 and HuR, which reduced the rate of cell death in paclitaxel resistant oral cancer cells. Specific inhibition of COX-2 by celecoxib, promoted apoptosis through activation of caspase-3 and cleavage of HuR in paclitaxel-resistant oral cancer cells, both in vitro and in vivo. In addition, oral cancer cells overexpressing cellular HuR increased the half-life of COX-2 mRNA, promoted COX-2 protein expression and exhibited enhanced tumor growth in vivo in comparison with cells expressing a cleavable form of HuR. Finally, our ribonucleoprotein immunoprecipitation and sequencing (RIP-seq) analyses of HuR in oral cancer cells treated with ionizing radiation (IR), determined that HuR cleavage product-1 (HuR-CP1) bound and promoted the expression of mRNAs encoding proteins involved in apoptosis. Our results indicated that, cellular non-cleavable HuR controls COX-2 mRNA expression and enzymatic activity. In addition, overexpressed COX-2 protein repressed the cleavage of caspase-3 and HuR to promote drug resistance and tumor growth. Altogether, our observations support the use of the COX-2 inhibitor celecoxib, in combination with paclitaxel, for the management of paclitaxel resistant oral cancer cells.

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Abbreviations

H&E:

Hematoxylin and eosin

HuR-CP1:

Hu Antigen-R cleavage product-1

HuR-D226A:

Non-cleavable isoform of HuR

RBP:

RNA-binding protein

ARE:

AU-rich elements

3′ UTR:

Untranslated region

HNSCC:

Head and neck squamous cell carcinoma

IR:

Ionizing radiation

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Acknowledgements

We acknowledge the funding source from the National Institutes of Health R01DE022776 and R01DE022776S1 to VP. We acknowledge the Center for Oral Health Research (L-COHR) which is supported by the NIH grant P30 GM103331.

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Authors and Affiliations

  1. Department of Oral Health Sciences and Center for Oral Health Research, College of Dental Medicine, Medical University of South Carolina, Charleston, SC, USA

    H Janakiraman, R P House, S Talwar & V Palanisamy

  2. Center for Genomics Medicine, Medical University of South Carolina, Charleston, SC, USA

    S M Courtney, E S Hazard & G Hardiman

  3. Department of Pathology, Medical University of South Carolina, Charleston, SC, USA

    S M Courtney

  4. Library Science and Informatics, Medical University of South Carolina, Charleston, SC, USA

    E S Hazard

  5. Departments of Medicine and Public Health, Medical University of South Carolina, Charleston, SC, USA

    G Hardiman

  6. Department of Surgery, Medical University of South Carolina, Charleston, SC, USA

    S Mehrotra

  7. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC, USA

    P H Howe, V Gangaraju & V Palanisamy

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Correspondence to V Palanisamy.

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Janakiraman, H., House, R., Talwar, S. et al. Repression of caspase-3 and RNA-binding protein HuR cleavage by cyclooxygenase-2 promotes drug resistance in oral squamous cell carcinoma. Oncogene 36, 3137–3148 (2017). https://doi.org/10.1038/onc.2016.451

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