Keratin14 mRNA expression in human pneumocytes during quiescence, repair and disease

PLoS One. 2017 Feb 15;12(2):e0172130. doi: 10.1371/journal.pone.0172130. eCollection 2017.

Abstract

The lung alveoli slowly self-renew pneumocytes, but their facultative regeneration capacity is rapidly efficient after an injury, so fibrosis infrequently occurs. We recently observed Keratin 14 (KRT14) expression during diffuse alveolar damage (DAD), but not in controls. We wonder if KRT14 may be a marker of pneumocyte transition from quiescence to regeneration. Quantitative PCR and Western blot analyses highlighted the presence of KRT14 (mRNA and protein) only in human lung samples with DAD or interstitial lung disease (ILD). In the exponentially growing cell lines A549 and H441, the mRNA and protein levels of KRT14 peaked at day one after cell seeding and decreased at day two, opposite to what observed for the proliferation marker E2F1. The inverse relation of KRT14 versus E2F1 expression holds true also for other proliferative markers, such as cyclin E1 and cyclin D1. Of interest, we also found that E2F1 silencing caused cell cycle arrest and increased KRT14 expression, whilst E2F1 stimulation induced cell cycle progression and decreased KRT14. KRT14 also increased in proliferative pneumocytes (HPAEpiC) just before transdifferentiation. Overall, our results suggest that KRT14 is a viable biomarker of pneumocyte activation, and repair/regeneration. The involvement of KRT14 in regenerative process may suggest a novel pharmaceutical target to accelerate lung repair.

MeSH terms

  • A549 Cells
  • Adolescent
  • Aged
  • Aged, 80 and over
  • Alveolar Epithelial Cells / cytology
  • Alveolar Epithelial Cells / metabolism
  • Cell Cycle Checkpoints
  • Cell Proliferation
  • Cell Transdifferentiation
  • Cells, Cultured
  • Cyclin D1 / metabolism
  • Cyclin E / metabolism
  • E2F1 Transcription Factor / antagonists & inhibitors
  • E2F1 Transcription Factor / genetics
  • E2F1 Transcription Factor / metabolism
  • Female
  • Humans
  • Keratin-14 / antagonists & inhibitors
  • Keratin-14 / genetics*
  • Keratin-14 / metabolism
  • Lung Diseases, Interstitial / metabolism
  • Lung Diseases, Interstitial / pathology*
  • Male
  • Middle Aged
  • Oncogene Proteins / metabolism
  • RNA Interference
  • RNA, Messenger / metabolism*
  • RNA, Small Interfering / metabolism
  • Transcriptome

Substances

  • CCND1 protein, human
  • CCNE1 protein, human
  • Cyclin E
  • E2F1 Transcription Factor
  • E2F1 protein, human
  • Keratin-14
  • Oncogene Proteins
  • RNA, Messenger
  • RNA, Small Interfering
  • Cyclin D1

Grants and funding

This work was in part supported by the “Beneficentia Stiftung” of Vaduz Liechtenstein, by the Italian Minister of Instruction, University and Research (MIUR), PRIN 2010-11, number 20109PLMH2 and by F.R.A., University of Trieste 2015.