Clusterin is a gene-specific target of microrna-21 in head and neck squamous cell carcinoma

Wojciech Mydlarz, Mamoru Uemura, Sun Ahn, Patrick Hennessey, Steven Chang, Semra Demokan, Wenyue Sun, Chunbo Shao, Justin Bishop, Julie Krosting, Elizabeth Mambo, William Westra, Patrick Ha, David Sidransky, Joseph Califano

Research output: Contribution to journalArticle

Abstract

Purpose: MicroRNA-21 (miRNA-21) has proto-oncogenic properties, although no miRNA-21-specific targets have been found in head and neck squamous cell carcinoma (HNSCC). Further study of miRNA-21 and its specific targets is essential to understanding HNSCC biology. Experimental Design: miRNA expression profiles of 10 HNSCCs and 10 normal mucosa samples were investigated using a custom miRNA microarray. Thirteen HNSCCs and five normal mucosa primary tissue specimens underwent mRNA expression microarray analysis. To identify miRNA-21 downstream targets, oral keratinocyte cells were subjected to microarray analysis after miRNA-21 transient transfection. miRNA and mRNA expression were validated by reverse transcription quantitative polymerase chain reaction (RTqPCR) in a separate cohort of 16 HNSCCs and 15 normal mucosal samples. Microarray and bioinformatics analyses were integrated to identify potential gene targets. In vitro assays looked at the function and interaction of miRNA-21 and its specific gene targets. Results: miRNA-21 was upregulated in HNSCCs and stimulated cell growth. Integrated analyses identified Clusterin (CLU) as a potential miRNA-21 gene target. CLU was downregulated after forced expression of miRNA-21 in normal and HNSCC cell lines. The activity of a luciferase construct containing the 30-untranslated region (UTR) of CLU was repressed by the ectopic expression of miRNA-21. CLU was also downregulated in primary HNSCCs and correlated with miRNA-21 overexpression. CLU variant 1 (CLU-1) was the predominant splice variant in HNSCCs and showed growth suppression function that was reversed by miRNA-21 overexpression. Conclusions: CLUis a specific, functional target ofoncogenic miRNA-21 inHNSCCs.CLU-1 isoformis the predominantgrowth-suppressivevarianttargetedbymiRNA-21.

Original languageEnglish (US)
Pages (from-to)868-877
Number of pages10
JournalClinical Cancer Research
Volume20
Issue number4
DOIs
StatePublished - 2014

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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    Mydlarz, W., Uemura, M., Ahn, S., Hennessey, P., Chang, S., Demokan, S., Sun, W., Shao, C., Bishop, J., Krosting, J., Mambo, E., Westra, W., Ha, P., Sidransky, D., & Califano, J. (2014). Clusterin is a gene-specific target of microrna-21 in head and neck squamous cell carcinoma. Clinical Cancer Research, 20(4), 868-877. https://doi.org/10.1158/1078-0432.CCR-13-2675