E2F6 protein levels modulate drug induced apoptosis in cardiomyocytes

Cell Signal. 2017 Dec:40:230-238. doi: 10.1016/j.cellsig.2017.09.022. Epub 2017 Sep 28.

Abstract

The E2F/Rb pathway regulates cell growth, differentiation, and death. In particular, E2F1 promotes apoptosis in all cells including those of the heart. E2F6, which represses E2F activity, was found to induce dilated cardiomyopathy in the absence of apoptosis in murine post-natal heart. Here we evaluate the anti-apoptotic potential of E2F6 in neonatal cardiomyocytes (NCM) from E2F6-Tg hearts which showed significantly less caspase-3 cleavage, a lower Bax/Bcl2 ratio, and improved cell viability in response to CoCl2 exposure. This correlated with a decrease in the pro-apoptotic E2F3 protein levels. In contrast, no difference in apoptotic markers or cell viability was observed in response to Doxorubicin (Dox) treatment between Wt and Tg-NCM. Dox caused a rapid and dramatic loss of the E2F6 protein in Tg-NCM within 6h and was undetectable after 12h. The level of e2f6 transcript was unchanged in Wt NCM, but was dramatically decreased in Tg cells in response to both Dox and CoCl2. This was related to an impact of the drugs on the α-myosin heavy chain promoter used to drive the E2F6 transgene. By comparison in HeLa, Dox induced apoptosis through upregulation of endogenous E2F1 involving post-transcriptional mechanisms, while E2F6 was down regulated with induction of the Checkpoint kinase-1 and proteasome degradation. These data imply that E2F6 serves to modulate E2F activity and protect cells including cardiomyocytes from apoptosis and improve survival. Strategies to modulate E2F6 levels may be therapeutically useful to mitigate cell death associated disorders.

Keywords: Cell survival; Dilated cardiomyopathy; Heart failure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Apoptosis / drug effects
  • Apoptosis / genetics*
  • Cardiomyopathy, Dilated
  • Caspase 3 / genetics
  • Cell Differentiation / genetics
  • Cell Proliferation / genetics
  • Doxorubicin / administration & dosage
  • E2F1 Transcription Factor / genetics*
  • E2F3 Transcription Factor / genetics
  • E2F6 Transcription Factor / genetics*
  • Gene Expression Regulation, Developmental / drug effects
  • HeLa Cells
  • Humans
  • Mice
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism*

Substances

  • E2F1 Transcription Factor
  • E2F3 Transcription Factor
  • E2F6 Transcription Factor
  • E2f1 protein, mouse
  • E2f3 protein, mouse
  • E2f6 protein, mouse
  • Doxorubicin
  • Casp3 protein, mouse
  • Caspase 3