Mu-opioid receptor activation promotes in vitro and in vivo tumor growth in head and neck squamous cell carcinoma

doi:10.1016/j.lfs.2021.119541

Life Sciences

Volume 278, 1 August 2021, 119541

https://doi.org/10.1016/j.lfs.2021.119541 Get rights and content

Highlights

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Mu-opioid receptor activation can drive cell growth and metastasis in MOR (+) HNSCC.
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DAMGO increases cell proliferation, colony formation, migration and invasion in MOR (+) HNSCC cells in vitro.
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DAMGO significantly increases the tumor volume compared to placebo.
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DAMGO decreases tumor growth and the mean time for tumor formation in an orthotopic HNSCC mouse model.

Abstract

Aims

In vitro and in vivo studies suggest that the mu-opioid receptor (MOR) is involved in tumorigenesis, and metastasis in cancer. In humans, the use of MOR agonists (opioids) is associated with head and neck squamous cell carcinoma (HNSCC) progression. In the present study, we aimed to examine the role of MOR activation in MOR (+) HNSCC.

Main methods

FaDu, MDA686Tu and UMSCC47 cell lines were used in in vitro and in vivo experiments. Cells and animals were treated with a highly selective MOR agonist DAMGO, [D-Ala (2), Me Phe (4), Glycol (5)]-enkephalin] or saline 0.9%.

Key findings

MOR activation significantly increased the proliferation, colony formation, invasion, and migration of FaDu and MDA6868Tu cells and promoted tumor growth in vivo.

Significance

These findings suggest that MOR is implicated in tumorigenesis of HNSCC. Overall, our findings identify that MOR could be used as a potential therapeutic target in patients with MOR (+) HNSCC.

Graphical abstract

Introduction

Head and neck cancers are the sixth most common malignancies worldwide (1). Despite recent advances in cancer treatment, the prognosis of patients with head and neck squamous cell carcinomas (HNSCC) has not significantly improved over the last years, especially in those patients with advanced disease (2). Patients with HNSCC can experience significant pain as the result of their cancers or the anticancer therapies (3). In those patients, the prescription of opioids remains as a main strategy for pain management (4).

The administration of opioids might be associated with worse oncological outcomes in patients with HNSCC. Our group demonstrated that the use of large dosages of opioids was associated with worse survival in patients undergoing surgery for laryngeal squamous cell carcinoma (5). The mu-opioid receptor (MOR) is the site of action of prescription opioids (6). MOR is upregulated in laryngeal carcinomas compared to normal tissues (7). New emerging data also suggest that tumoral MOR expression is an independent predictor of oncological outcomes in head and neck cancers. A study by Zhang et al. showed that a high level of expression of MOR was associated with shorter recurrence-free and overall survival in patients with laryngeal SCC (8).

Several laboratory studies have investigated the role of MOR in different malignancies. In lung and breast cancer cells, MOR agonists such as DAMGO [D-Ala (2), Me Phe (4), Glycol (5)]-enkephalin] stimulate proliferation, survival, and migration (9,10). While an in vitro study by Zagon's group demonstrated that DAMGO (10⁻⁶ M for seven days) promoted apoptosis in CAL-27 HNSCC cells not expressing MOR (11). The authors speculated that the apoptotic effects of DAMGO were mediated by their “synthetic nature” or independent to its activity on MOR (12).

This laboratory investigation aimed to study the role of MOR activation in tumorigenesis and metastasis in HNSCC. We tested the hypothesis that MOR stimulation promoted, in vitro, proliferation, survival, migration, and invasion of HNSCC cells and, in vivo, induced tumor growth.

Section snippets

Cell lines and cell culture conditions

The human HNSCC cell line FaDu was purchased from the American Type Culture Collection (Manassas, VA). MDA686Tu and UMSCC47 cells were obtained from the laboratory of Dr. Jeffrey N. Myers, The University of Texas M.D. Anderson Cancer Center. All cell lines used in this study were authenticated as mycoplasma free. Mycoplasma testing was performed using The MycoAlert® Mycoplasma Detection Kit (Lonza). Cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum

Expression and activity of MOR in HNSCC cells

RNA sequencing data indicated that FaDu and MDA686Tu cells expressed OPRM1 gene while UMSCC47 cells did not express the gene. Western blot analyses confirmed RNA sequencing data and revealed that MOR protein expression levels were significantly higher in FaDu and MDA686Tu cell lines than in UMSCC47 cells (Fig. 1A-1B). In general, opioids are known to inhibit adenylyl cyclase activity and decrease cAMP formation (18). However, depending on the experimental conditions such as cell type and

Discussion

The impact of MOR activation in HNSCC is not yet fully understood. In this study, we observed that MOR activation via DAMGO enhanced tumorigenesis and metastatic cell behaviors in HNSCC cells expressing the receptor. Our pharmacological and genetic studies blocking and silencing MOR confirm that the observed effects of DAMGO were mediated by activation of the receptor. We also found that a low-dose of DAMGO accelerated tumor growth in vivo. Based on previous studies, there is a common

Conclusion

In summary, our results suggest that MOR activation can play a specific role in cancer progression and tumorigenesis and could be considered as a target for HNSCC therapy. More research is needed to elucidate the mechanisms underneath and whether clinically available antagonists of the receptor can be used introduced clinically in combination with adjuvant therapies.

Author contributions

A. Gorur: Investigation, Validation, Resources, Methodology, Writing - Original Draft, Writing - Review & Editing. M. Patiño: Investigation, Writing - Review & Editing. H. Takahashi: Investigation, Writing - Review & Editing, G. Corrales: Investigation, Writing - Review & Editing. C.R. Pickering: Investigation, Writing - Review & Editing. F.O. Gleber-Netto: Investigation, Writing - Review & Editing. J.N. Myers: Writing - Review & Editing, Methodology, Supervision. J. Cata: Conceptualization,

Acknowledgements

We would like to thank The University of Texas MD Anderson Cancer Center for supporting the grant and all the studies in the manuscript.

Funding

This work was supported by the University of Texas MD Anderson Cancer Center start-up research project [Grant number, 00001816-RN00].

Data availability statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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