Repression of p63 and induction of EMT by mutant Ras in mammary epithelial cells

Kathryn E Yoh; Kausik Regunath; Asja Guzman; Seung-Min Lee; Neil T Pfister; Olutosin Akanni; Laura J Kaufman; Carol Prives; Ron Prywes

doi:10.1073/pnas.1613417113

Repression of p63 and induction of EMT by mutant Ras in mammary epithelial cells

Proc Natl Acad Sci U S A. 2016 Oct 11;113(41):E6107-E6116. doi: 10.1073/pnas.1613417113. Epub 2016 Sep 28.

Authors

Kathryn E Yoh¹, Kausik Regunath¹, Asja Guzman², Seung-Min Lee³, Neil T Pfister¹, Olutosin Akanni¹, Laura J Kaufman², Carol Prives⁴, Ron Prywes⁴

Affiliations

¹ Department of Biological Sciences, Columbia University, New York, NY 10027.
² Department of Chemistry, Columbia University, New York, NY 10027.
³ Department of Food and Nutritional Sciences, College of Human Ecology, Yonsei University, Seoul 03722, South Korea.
⁴ Department of Biological Sciences, Columbia University, New York, NY 10027; [email protected] [email protected].

Abstract

The p53-related transcription factor p63 is required for maintenance of epithelial cell differentiation. We found that activated forms of the Harvey Rat Sarcoma Virus GTPase (H-RAS) and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) oncogenes strongly repress expression of ∆Np63α, the predominant p63 isoform in basal mammary epithelial cells. This regulation occurs at the transcriptional level, and a short region of the ∆Np63 promoter is sufficient for repression induced by H-RasV12. The suppression of ∆Np63α expression by these oncogenes concomitantly leads to an epithelial-to-mesenchymal transition (EMT). In addition, the depletion of ∆Np63α alone is sufficient to induce EMT. Both H-RasV12 expression and ∆Np63α depletion induce individual cell invasion in a 3D collagen gel in vitro system, thereby demonstrating how Ras can drive the mammary epithelial cell state toward greater invasive ability. Together, these results suggest a pathway by which RAS and PIK3CA oncogenes induce EMT through regulation of ∆Np63α.

Keywords: H-Ras; breast cancer; epithelial mesenchymal transition; p63; transcriptional repression.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Base Sequence
Binding Sites
Cell Line, Tumor
Cell Transformation, Neoplastic / genetics
Cell Transformation, Neoplastic / metabolism
Epithelial Cells / metabolism*
Epithelial-Mesenchymal Transition / genetics*
Gene Expression Regulation*
Genes, Reporter
Humans
Models, Biological
Mutation*
Phosphatidylinositol 3-Kinases / metabolism
Promoter Regions, Genetic
Protein Binding
Proto-Oncogene Proteins c-akt / metabolism
RNA Interference
RNA, Small Interfering / genetics
Sequence Deletion
Signal Transduction
Transcription Factors / chemistry
Transcription Factors / genetics*
Transcription Factors / metabolism
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism
Tumor Suppressor Proteins / chemistry
Tumor Suppressor Proteins / genetics*
Tumor Suppressor Proteins / metabolism
ras Proteins / genetics*

Substances

RNA, Small Interfering
TP63 protein, human
Transcription Factors
Tumor Suppressor Protein p53
Tumor Suppressor Proteins
Phosphatidylinositol 3-Kinases
Proto-Oncogene Proteins c-akt
ras Proteins

Grants and funding

P01 CA087497/CA/NCI NIH HHS/United States