The cell cycle regulator ecdysoneless cooperates with H-ras to promote oncogenic transformation of human mammary epithelial cells

Aditya Bele, Sameer Mirza, Ying Zhang, Riyaz Ahmad Mir, Simon Lin, Jun Hyun Kim, Channabasavaiah B Gurumurthy, William West, Fang Qiu, Hamid Band, Vimla Band

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

The mammalian ortholog of Drosophila ecdysoneless (Ecd) gene product regulates Rb-E2F interaction and is required for cell cycle progression. Ecd is overexpressed in breast cancer and its overexpression predicts shorter survival in patients with ErbB2-positive tumors. Here, we demonstrate Ecd knock down (KD) in human mammary epithelial cells (hMECs) induces growth arrest, similar to the impact of Ecd Knock out (KO) in mouse embryonic fibroblasts. Furthermore, whole-genome mRNA expression analysis of control vs. Ecd KD in hMECs demonstrated that several of the top 40 genes that were down-regulated were E2F target genes. To address the role of Ecd in mammary oncogenesis, we overexpressed Ecd and/or mutant H-Ras in hTERT-immortalized hMECs. Cell cycle analyses revealed hMECs overexpressing EcdCRas showed incomplete arrest in G1 phase upon growth factor deprivation, and more rapid cell cycle progression in growth factor-containing medium. Analyses of cell migration, invasion, acinar structures in 3-D Matrigel and anchorage-independent growth demonstrated that EcdCRas-overexpressing cells exhibit substantially more dramatic transformed phenotype as compared to cells expressing vector, Ras or Ecd. Under conditions of nutrient deprivation, EcdCRas-overexpressing hMECs exhibited better survival, with substantial upregulation of the autophagy marker LC3 both at the mRNA and protein levels. Significantly, while hMECs expressing Ecd or mutant Ras alone did not form tumors in NOD/SCID mice, EcdCRas-overexpressing hMECs formed tumors, clearly demonstrating oncogenic cooperation between Ecd and mutant Ras. Collectively, we demonstrate an important co-oncogenic role of Ecd in the progression of mammary oncogenesis through promoting cell survival.

Original languageEnglish (US)
Pages (from-to)990-1000
Number of pages11
JournalCell Cycle
Volume14
Issue number7
DOIs
StatePublished - Apr 1 2015

Keywords

  • Autophagy
  • Breast cancer
  • Cell cycle
  • Ecdysoneless
  • Ras

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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