Investigating miR-9 as a mediator in laryngeal cancer health disparities

Christina Gobin; Samuel Inkabi; Chayil C Lattimore; Tongjun Gu; James N Menefee; Mayrangela Rodriguez; Heather Kates; Christopher Fields; Tengfei Bian; Natalie Silver; Chengguo Xing; Clayton Yates; Rolf Renne; Mingyi Xie; Kristianna M Fredenburg

doi:10.3389/fonc.2023.1096882

Investigating miR-9 as a mediator in laryngeal cancer health disparities

Front Oncol. 2023 Apr 4:13:1096882. doi: 10.3389/fonc.2023.1096882. eCollection 2023.

Authors

Christina Gobin¹, Samuel Inkabi², Chayil C Lattimore¹, Tongjun Gu³, James N Menefee¹, Mayrangela Rodriguez¹, Heather Kates¹, Christopher Fields⁴, Tengfei Bian⁵, Natalie Silver⁶, Chengguo Xing⁵, Clayton Yates^{7

8

9}, Rolf Renne¹⁰, Mingyi Xie¹¹, Kristianna M Fredenburg¹

Affiliations

¹ Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, Gainesville, FL, United States.
² College of Graduate Health Studies, A.T. Still University, Kirksville, MO, United States.
³ Interdisciplinary Center for Biotechnology Research Bioinformatics Core Facility, University of Florida, Gainesville, FL, United States.
⁴ Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, United States.
⁵ Department of Medicinal Chemistry, University of Florida, Gainesville, FL, United States.
⁶ Head and Neck Institute/Lerner Research Institute, Cleveland Clinic, Cleveland, OH, United States.
⁷ Department of Pathology, Johns Hopkins School of Medicine, Baltimore, MD, United States.
⁸ Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
⁹ Department of Urology, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
¹⁰ Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL, United States.
¹¹ Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL, United States.

Abstract

Background: For several decades, Black patients have carried a higher burden of laryngeal cancer among all races. Even when accounting for sociodemographics, a disparity remains. Differentially expressed microRNAs have been linked to racially disparate clinical outcomes in breast and prostate cancers, yet an association in laryngeal cancer has not been addressed. In this study, we present our computational analysis of differentially expressed miRNAs in Black compared with White laryngeal cancer and further validate microRNA-9-5p (miR-9-5p) as a potential mediator of cancer phenotype and chemoresistance.

Methods: Bioinformatic analysis of 111 (92 Whites, 19 Black) laryngeal squamous cell carcinoma (LSCC) specimens from the TCGA revealed miRNAs were significantly differentially expressed in Black compared with White LSCC. We focused on miR-9-5 p which had a significant 4-fold lower expression in Black compared with White LSCC (p<0.05). After transient transfection with either miR-9 mimic or inhibitor in cell lines derived from Black (UM-SCC-12) or White LSCC patients (UM-SCC-10A), cellular migration and cell proliferation was assessed. Alterations in cisplatin sensitivity was evaluated in transient transfected cells via IC50 analysis. qPCR was performed on transfected cells to evaluate miR-9 targets and chemoresistance predictors, ABCC1 and MAP1B.

Results: Northern blot analysis revealed mature miR-9-5p was inherently lower in cell line UM-SCC-12 compared with UM-SCC-10A. UM -SCC-12 had baseline increase in cellular migration (p < 0.01), proliferation (p < 0.0001) and chemosensitivity (p < 0.01) compared to UM-SCC-10A. Increasing miR-9 in UM-SCC-12 cells resulted in decreased cellular migration (p < 0.05), decreased proliferation (p < 0.0001) and increased sensitivity to cisplatin (p < 0.001). Reducing miR-9 in UM-SCC-10A cells resulted in increased cellular migration (p < 0.05), increased proliferation (p < 0.05) and decreased sensitivity to cisplatin (p < 0.01). A significant inverse relationship in ABCC1 and MAP1B gene expression was observed when miR-9 levels were transiently elevated or reduced in either UM-SCC-12 or UM-SCC-10A cell lines, respectively, suggesting modulation by miR-9.

Conclusion: Collectively, these studies introduce differential miRNA expression in LSCC cancer health disparities and propose a role for low miR-9-5p as a mediator in LSCC tumorigenesis and chemoresistance.

Keywords: ABCC1; MAP1B; cancer health disparities; head and neck cancer; laryngeal squamous cell carcinoma; miR-9.

Abstract

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