miR-375 Regulates Invasion-Related Proteins Vimentin and L-Plastin

Lizandra Jimenez, Jihyeon Lim, Berta Burd, Thomas M. Harris, Thomas J. Ow, Nicole Kawachi, Thomas J. Belbin, Ruth Angeletti, Michael B. Prystowsky, Geoffrey J. Childs, Jeffrey E. Segall

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Invasion is a hallmark of advanced head and neck squamous cell carcinoma (HNSCC). We previously determined that low relative miR-375 expression was associated with poor patient prognosis. HNSCC cells with increased miR-375 expression have lower invasive properties and impaired invadopodium activity. Using stable isotope labeling with amino acids in cell culture and reverse-phase liquid chromatography mass spectrometry, we assessed the impact of miR-375 expression on protein levels in UM-SCC-1 cells. Increased miR-375 expression was associated with down-regulation of proteins involved in cellular assembly and organization, death and survival, and movement. Two invasion-associated proteins, vimentin and L-plastin, were strongly down-regulated by miR-375. Luciferase reporter assays demonstrated that high miR-375 expression reduced vimentin promoter activity, suggesting that vimentin is an indirect target of miR-375. Runt-related transcription factor 1 (RUNX1) is a potential miR-375 direct target, and its knockdown reduced vimentin and L-plastin expression. Data in The Cancer Genome Atlas HNSCC database showed a significant inverse correlation between miR-375 expression and RUNX1, vimentin, and L-plastin RNA expression. These clinical correlations validate our in vitro model findings and support a mechanism in which miR-375 suppresses RUNX1 levels, resulting in reduced vimentin and L-plastin expression. Furthermore, knockdown of RUNX1, L-plastin, and vimentin resulted in significant reductions in cell invasion in vitro, indicating the functional significance of miR-375 regulation of specific proteins involved in HNSCC invasion.

Original languageEnglish (US)
Pages (from-to)1523-1536
Number of pages14
JournalAmerican Journal of Pathology
Volume187
Issue number7
DOIs
StatePublished - Jul 1 2017

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Vimentin
Proteins
Core Binding Factor Alpha 2 Subunit
Isotope Labeling
Atlases
Reverse-Phase Chromatography
plastin
Luciferases
Mass Spectrometry
Down-Regulation
Cell Culture Techniques
Genome
Databases
RNA
Amino Acids
Survival
Carcinoma, squamous cell of head and neck
Neoplasms

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

miR-375 Regulates Invasion-Related Proteins Vimentin and L-Plastin. / Jimenez, Lizandra; Lim, Jihyeon; Burd, Berta; Harris, Thomas M.; Ow, Thomas J.; Kawachi, Nicole; Belbin, Thomas J.; Angeletti, Ruth; Prystowsky, Michael B.; Childs, Geoffrey J.; Segall, Jeffrey E.

In: American Journal of Pathology, Vol. 187, No. 7, 01.07.2017, p. 1523-1536.

Research output: Contribution to journalArticle

Jimenez, Lizandra ; Lim, Jihyeon ; Burd, Berta ; Harris, Thomas M. ; Ow, Thomas J. ; Kawachi, Nicole ; Belbin, Thomas J. ; Angeletti, Ruth ; Prystowsky, Michael B. ; Childs, Geoffrey J. ; Segall, Jeffrey E. / miR-375 Regulates Invasion-Related Proteins Vimentin and L-Plastin. In: American Journal of Pathology. 2017 ; Vol. 187, No. 7. pp. 1523-1536.
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abstract = "Invasion is a hallmark of advanced head and neck squamous cell carcinoma (HNSCC). We previously determined that low relative miR-375 expression was associated with poor patient prognosis. HNSCC cells with increased miR-375 expression have lower invasive properties and impaired invadopodium activity. Using stable isotope labeling with amino acids in cell culture and reverse-phase liquid chromatography mass spectrometry, we assessed the impact of miR-375 expression on protein levels in UM-SCC-1 cells. Increased miR-375 expression was associated with down-regulation of proteins involved in cellular assembly and organization, death and survival, and movement. Two invasion-associated proteins, vimentin and L-plastin, were strongly down-regulated by miR-375. Luciferase reporter assays demonstrated that high miR-375 expression reduced vimentin promoter activity, suggesting that vimentin is an indirect target of miR-375. Runt-related transcription factor 1 (RUNX1) is a potential miR-375 direct target, and its knockdown reduced vimentin and L-plastin expression. Data in The Cancer Genome Atlas HNSCC database showed a significant inverse correlation between miR-375 expression and RUNX1, vimentin, and L-plastin RNA expression. These clinical correlations validate our in vitro model findings and support a mechanism in which miR-375 suppresses RUNX1 levels, resulting in reduced vimentin and L-plastin expression. Furthermore, knockdown of RUNX1, L-plastin, and vimentin resulted in significant reductions in cell invasion in vitro, indicating the functional significance of miR-375 regulation of specific proteins involved in HNSCC invasion.",
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AU - Lim, Jihyeon

AU - Burd, Berta

AU - Harris, Thomas M.

AU - Ow, Thomas J.

AU - Kawachi, Nicole

AU - Belbin, Thomas J.

AU - Angeletti, Ruth

AU - Prystowsky, Michael B.

AU - Childs, Geoffrey J.

AU - Segall, Jeffrey E.

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AB - Invasion is a hallmark of advanced head and neck squamous cell carcinoma (HNSCC). We previously determined that low relative miR-375 expression was associated with poor patient prognosis. HNSCC cells with increased miR-375 expression have lower invasive properties and impaired invadopodium activity. Using stable isotope labeling with amino acids in cell culture and reverse-phase liquid chromatography mass spectrometry, we assessed the impact of miR-375 expression on protein levels in UM-SCC-1 cells. Increased miR-375 expression was associated with down-regulation of proteins involved in cellular assembly and organization, death and survival, and movement. Two invasion-associated proteins, vimentin and L-plastin, were strongly down-regulated by miR-375. Luciferase reporter assays demonstrated that high miR-375 expression reduced vimentin promoter activity, suggesting that vimentin is an indirect target of miR-375. Runt-related transcription factor 1 (RUNX1) is a potential miR-375 direct target, and its knockdown reduced vimentin and L-plastin expression. Data in The Cancer Genome Atlas HNSCC database showed a significant inverse correlation between miR-375 expression and RUNX1, vimentin, and L-plastin RNA expression. These clinical correlations validate our in vitro model findings and support a mechanism in which miR-375 suppresses RUNX1 levels, resulting in reduced vimentin and L-plastin expression. Furthermore, knockdown of RUNX1, L-plastin, and vimentin resulted in significant reductions in cell invasion in vitro, indicating the functional significance of miR-375 regulation of specific proteins involved in HNSCC invasion.

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