miR-22 and miR-205 Drive Tumor Aggressiveness of Mucoepidermoid Carcinomas of Salivary Glands

Erika Naakka; Mateus Camargo Barros-Filho; Shady Adnan-Awad; Ahmed Al-Samadi; Fábio Albuquerque Marchi; Hellen Kuasne; Katja Korelin; Ilida Suleymanova; Amy Louise Brown; Cristovam Scapulatempo-Neto; Silvia Vanessa Lourenço; Rogério Moraes Castilho; Luiz Paulo Kowalski; Antti Mäkitie; Vera Cavalcanti Araújo; Ilmo Leivo; Silvia Regina Rogatto; Tuula Salo; Fabricio Passador-Santos

doi:10.3389/fonc.2021.786150

miR-22 and miR-205 Drive Tumor Aggressiveness of Mucoepidermoid Carcinomas of Salivary Glands

Front Oncol. 2022 Feb 9:11:786150. doi: 10.3389/fonc.2021.786150. eCollection 2021.

Authors

Erika Naakka^{1

2}, Mateus Camargo Barros-Filho³, Shady Adnan-Awad^{2

4}, Ahmed Al-Samadi^{1

2}, Fábio Albuquerque Marchi³, Hellen Kuasne³, Katja Korelin^{1

2}, Ilida Suleymanova^{1

2}, Amy Louise Brown⁵, Cristovam Scapulatempo-Neto⁶, Silvia Vanessa Lourenço^{7

8}, Rogério Moraes Castilho⁹, Luiz Paulo Kowalski^{10

11}, Antti Mäkitie^{12

13

14}, Vera Cavalcanti Araújo⁵, Ilmo Leivo¹⁵, Silvia Regina Rogatto^{16

17}, Tuula Salo^{1

2

18

19

20}, Fabricio Passador-Santos⁵

Affiliations

¹ Department of Oral and Maxillofacial Diseases, University of Helsinki, Helsinki, Finland.
² Translational Immunology Research Program (TRIMM), University of Helsinki, Helsinki, Finland.
³ Centro Internacional de Pesquisa (CIPE) - A.C.Camargo Cancer Center, São Paulo, Brazil.
⁴ Hematology Research Unit, Department of Clinical Chemistry and Hematology, University of Helsinki, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland.
⁵ Department of Oral Pathology, Faculdade São Leopoldo Mandic, Campinas, Brazil.
⁶ Molecular Oncology Research Center, Barretos, and Diagnósticos da América (DASA), Barueri, Brazil.
⁷ Department of Pathology, A.C.Camargo Cancer Center, São Paulo, Brazil.
⁸ Department of General Pathology, Dental School, University of São Paulo, São Paulo, Brazil.
⁹ Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, MI, United States.
¹⁰ Department of Head and Neck Surgery and Otorhinolaryngology, A.C.Camargo Cancer Center, São Paulo, Brazil.
¹¹ Department of Head and Neck Surgery, University of Sao Paulo Medical School, São Paulo, Brazil.
¹² Department of Otorhinolaryngology - Head and Neck Surgery, University of Helsinki and Helsinki University Hospital, Helsinki, Finland.
¹³ Research Program in Systems Oncology, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
¹⁴ Division of Ear, Nose and Throat Diseases, Department of Clinical Sciences, Intervention and Technology, Karolinska Institute and Karolinska Hospital, Stockholm, Sweden.
¹⁵ Institute of Biomedicine, Pathology, University of Turku and Turku University Hospital, Turku, Finland.
¹⁶ Department of Clinical Genetics, University Hospital of Southern Denmark, Vejle, Denmark.
¹⁷ Institute of Regional Health Research, University of Southern Denmark, Odense, Denmark.
¹⁸ Department of Pathology, Helsinki University Hospital, Helsinki, Finland.
¹⁹ Cancer and Translational Medicine Research Unit, University of Oulu, Oulu, Finland.
²⁰ Medical Research Center, Oulu University Hospital, Oulu, Finland.

Abstract

Objectives: To integrate mRNA and miRNA expression profiles of mucoepidermoid carcinomas (MECs) and normal salivary gland (NSGs) tissue samples and identify potential drivers.

Material and methods: Gene and miRNA expression arrays were performed in 35 MECs and six NSGs.

Results: We found 46 differentially expressed (DE) miRNAs and 3,162 DE mRNAs. Supervised hierarchical clustering analysis of the DE transcripts revealed two clusters in both miRNA and mRNA profiles, which distinguished MEC from NSG samples. The integrative miRNA-mRNA analysis revealed a network comprising 696 negatively correlated interactions (44 miRNAs and 444 mRNAs) involving cell signaling, cell cycle, and cancer-related pathways. Increased expression levels of miR-205-5p and miR-224-5p and decreased expression levels of miR-139-3p, miR-145-3p, miR-148a-3p, miR-186-5p, miR-338-3p, miR-363-3p, and miR-4324 were significantly related to worse overall survival in MEC patients. Two overexpressed miRNAs in MEC (miR-22 and miR-205) were selected for inhibition by the CRISPR-Cas9 method. Cell viability, migration, and invasion assays were performed using an intermediate grade MEC cell line. Knockout of miR-205 reduced cell viability and enhanced ZEB2 expression, while miR-22 knockout reduced cell migration and invasion and enhanced ESR1 expression. Our results indicate a distinct transcriptomic profile of MEC compared to NSG, and the integrative analysis highlighted miRNA-mRNA interactions involving cancer-related pathways, including PTEN and PI3K/AKT.

Conclusion: The in vitro functional studies revealed that miR-22 and miR-205 deficiencies reduced the viability, migration, and invasion of the MEC cells suggesting they are potential oncogenic drivers in MEC.

Keywords: head and neck cancer; miR205; miR22; microRNA; mucoepidermoid carcinoma; oral cancer; salivary gland tumor; transcriptomic analysis.

Copyright © 2022 Naakka, Barros-Filho, Adnan-Awad, Al-Samadi, Marchi, Kuasne, Korelin, Suleymanova, Brown, Scapulatempo-Neto, Lourenço, Castilho, Kowalski, Mäkitie, Araújo, Leivo, Rogatto, Salo and Passador-Santos.