title: Alcohol-dysregulated miR-30a and miR-934 in head and neck squamous cell carcinoma.
abstract: BACKGROUND
  Alcohol consumption is a well-established risk factor for head and neck squamous cell carcinoma (HNSCC); however, the molecular mechanisms by which alcohol promotes HNSCC pathogenesis and progression remain poorly understood. Our study sought to identify microRNAs that are dysregulated in alcohol-associated HNSCC and investigate their contribution to the malignant phenotype.
  
  
  METHODS
  Using RNA-sequencing data from 136 HNSCC patients, we compared the expression levels of 1,046 microRNAs between drinking and non-drinking cohorts. Dysregulated microRNAs were verified by qRT-PCR in normal oral keratinocytes treated with biologically relevant doses of ethanol and acetaldehyde. The most promising microRNA candidates were investigated for their effects on cellular proliferation and invasion, sensitivity to cisplatin, and expression of cancer stem cell genes. Finally, putative target genes were identified and evaluated in vitro to further establish roles for these miRNAs in alcohol-associated HNSCC.
  
  
  RESULTS
  From RNA-sequencing analysis we identified 8 miRNAs to be significantly upregulated in alcohol-associated HNSCCs. qRT-PCR experiments determined that among these candidates, miR-30a and miR-934 were the most highly upregulated in vitro by alcohol and acetaldehyde. Overexpression of miR-30a and miR-934 in normal and HNSCC cell lines produced up to a 2-fold increase in cellular proliferation, as well as induction of the anti-apoptotic gene BCL-2. Upon inhibition of these miRNAs, HNSCC cell lines exhibited increased sensitivity to cisplatin and reduced matrigel invasion. miRNA knockdown also indicated direct targeting of several tumor suppressor genes by miR-30a and miR-934.
  
  
  CONCLUSIONS
  Alcohol induces the dysregulation of miR-30a and miR-934, which may play crucial roles in HNSCC pathogenesis and progression. Future investigation of the alcohol-mediated pathways effecting these transformations will prove valuable for furthering the understanding and treatment of alcohol-associated HNSCC.
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. masaad@ucsd.edu.
authors.forename: Maarouf A
authors.initials: MA
authors.lastname: Saad
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. szkuo@ucsd.edu.
authors.forename: Selena Z
authors.initials: SZ
authors.lastname: Kuo
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. elham.rahimy@yale.edu.
authors.forename: Elham
authors.initials: E
authors.lastname: Rahimy
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. azou@ucsd.edu.
authors.forename: Angela E
authors.initials: AE
authors.lastname: Zou
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. akorrapa@ucsd.edu.
authors.forename: Avinaash
authors.initials: A
authors.lastname: Korrapati
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. mrahimy@ucsd.edu.
authors.forename: Mehran
authors.initials: M
authors.lastname: Rahimy
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. elkim@ucsd.edu.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Kim
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. h3zheng@ucsd.edu.
authors.forename: Hao
authors.initials: H
authors.lastname: Zheng
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. michael.yu@my.rfums.org.
authors.forename: Michael Andrew
authors.initials: MA
authors.lastname: Yu
authors.affiliation: Department of Pathology, Veterans Administration Health Care System, San Diego, CA, USA. jessica.wang-rodriguez@va.gov.|Department of Pathology, University of California, San Diego, CA, USA. jessica.wang-rodriguez@va.gov.
authors.forename: Jessica
authors.initials: J
authors.lastname: Wang-Rodriguez
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California, La Jolla, San Diego, CA, USA. wongkeko@ucsd.edu.
authors.forename: Weg M
authors.initials: WM
authors.lastname: Ongkeko
mesh_terms: D000079:Acetaldehyde / Q000633:toxicity; D000328:Adult; D000438:Alcohols / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D000431:Ethanol / Q000633:toxicity; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics*; D006258:Head and Neck Neoplasms / Q000235:genetics*; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077195:Squamous Cell Carcinoma of Head and Neck
chemical_list: D000438:Alcohols; C546086:MIRN30b microRNA, human; D035683:MicroRNAs; D000431:Ethanol; D000079:Acetaldehyde