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 Citations (Scopus)

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

Fingerprint

Cell Cycle
Breast
Epithelial Cells
Intercellular Signaling Peptides and Proteins
Carcinogenesis
Neoplasms
Retinoblastoma Protein
Messenger RNA
Inbred NOD Mouse
SCID Mice
Survival
Autophagy
G1 Phase
Growth
Knockout Mice
Genes
Drosophila
Cell Movement
Cell Survival
Up-Regulation

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

The cell cycle regulator ecdysoneless cooperates with H-ras to promote oncogenic transformation of human mammary epithelial cells. / Bele, Aditya; Mirza, Sameer; Zhang, Ying; Mir, Riyaz Ahmad; Lin, Simon; Kim, Jun Hyun; Gurumurthy, Channabasavaiah B; West, William; Qiu, Fang; Band, Hamid; Band, Vimla.

In: Cell Cycle, Vol. 14, No. 7, 01.04.2015, p. 990-1000.

Research output: Contribution to journalArticle

Bele, Aditya ; Mirza, Sameer ; Zhang, Ying ; Mir, Riyaz Ahmad ; Lin, Simon ; Kim, Jun Hyun ; Gurumurthy, Channabasavaiah B ; West, William ; Qiu, Fang ; Band, Hamid ; Band, Vimla. / The cell cycle regulator ecdysoneless cooperates with H-ras to promote oncogenic transformation of human mammary epithelial cells. In: Cell Cycle. 2015 ; Vol. 14, No. 7. pp. 990-1000.
@article{6ce2598325c2468597e349fc6e97fbd8,
title = "The cell cycle regulator ecdysoneless cooperates with H-ras to promote oncogenic transformation of human mammary epithelial cells",
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.",
keywords = "Autophagy, Breast cancer, Cell cycle, Ecdysoneless, Ras",
author = "Aditya Bele and Sameer Mirza and Ying Zhang and Mir, {Riyaz Ahmad} and Simon Lin and Kim, {Jun Hyun} and Gurumurthy, {Channabasavaiah B} and William West and Fang Qiu and Hamid Band and Vimla Band",
year = "2015",
month = "4",
day = "1",
doi = "10.1080/15384101.2015.1006982",
language = "English (US)",
volume = "14",
pages = "990--1000",
journal = "Cell Cycle",
issn = "1538-4101",
publisher = "Landes Bioscience",
number = "7",

}

TY - JOUR

T1 - The cell cycle regulator ecdysoneless cooperates with H-ras to promote oncogenic transformation of human mammary epithelial cells

AU - Bele, Aditya

AU - Mirza, Sameer

AU - Zhang, Ying

AU - Mir, Riyaz Ahmad

AU - Lin, Simon

AU - Kim, Jun Hyun

AU - Gurumurthy, Channabasavaiah B

AU - West, William

AU - Qiu, Fang

AU - Band, Hamid

AU - Band, Vimla

PY - 2015/4/1

Y1 - 2015/4/1

N2 - 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.

AB - 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.

KW - Autophagy

KW - Breast cancer

KW - Cell cycle

KW - Ecdysoneless

KW - Ras

UR - http://www.scopus.com/inward/record.url?scp=84928112260&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84928112260&partnerID=8YFLogxK

U2 - 10.1080/15384101.2015.1006982

DO - 10.1080/15384101.2015.1006982

M3 - Article

VL - 14

SP - 990

EP - 1000

JO - Cell Cycle

JF - Cell Cycle

SN - 1538-4101

IS - 7

ER -