Altered regulation of PDK4 expression promotes antiestrogen resistance in human breast cancer cells

William Walter, Jennifer Thomalla, Josh Bruhn, Dedra H. Fagan, Cheryl Zehowski, Douglas Yee, Andrew J Skildum

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Acquired or de novo resistance to the selective estrogen receptor modulators tamoxifen and fulvestrant (ICI) is a major barrier to successful treatment of breast cancer. Gene expression patterns in tamoxifen resistant (TamR-MCF-7) cells were compared to their parental cells (MCF-7L) to identify an aberrantly regulated metabolic pathway. TamR-MCF-7 cells are cross resistant to ICI and doxorubicin, and have increased mitochondrial DNA. A small subset of genes had altered expression in TamR-MCF-7 relative to MCF-7L cells. One of the genes, pyruvate dehydrogenase kinase-4 (PDK4), phosphorylates pyruvate dehydrogenase (PDH). PDK4 expression was elevated in TamR-MCF-7 cells; this result was also observed in a second model of acquired antiestrogen resistance. PDK4 expression is controlled in part by glucocorticoid response elements in the PDK4 gene promoter. In MCF-7L cells, PDK4 mRNA expression was insensitive to glucocorticoid receptor agonists, while dexamethasone dramatically increased PDK4 expression in TamR-MCF-7 cells. Using siRNA to knock down PDK4 expression increased TamR-MCF-7 sensitivity to ICI; in contrast adapting cells to growth in glucose depleted media did not affect ICI sensitivity. Despite TamR-MCF-7 cells high levels of PDK4 mRNA relative to MCF-7L, TamR-MCF-7 cells have increased PDH activity. Wild type MCF-7 cells are reported to be heterozygous for a G to A mutation that results in a substitution of threonine for alanine near PDK4′s catalytic site. We found loss of heterozygosity in TamR-MCF-7 cells; TamR-MCF-7 are homozygous for the wild type allele. These data support a role for altered regulation of PDH by PDK4 and altered substrate utilization in the development of drug resistance in human breast cancer cells.

Original languageEnglish (US)
Article number689
Pages (from-to)1-13
Number of pages13
JournalSpringerPlus
Volume4
Issue number1
DOIs
StatePublished - Dec 1 2015

Fingerprint

Estrogen Receptor Modulators
MCF-7 Cells
Breast Neoplasms
Pyruvic Acid
Oxidoreductases
Tamoxifen
Genes
Selective Estrogen Receptor Modulators
pyruvate dehydrogenase kinase 4
Messenger RNA
Loss of Heterozygosity
Glucocorticoid Receptors
Response Elements
Threonine
Metabolic Networks and Pathways
Mitochondrial DNA
Drug Resistance
Alanine
Doxorubicin
Dexamethasone

Keywords

  • Antiestrogen resistance
  • Breast cancer
  • Glucose
  • Metabolism
  • Pyruvate dehydrogenase kinase-isoform 4 (PDK4)

Cite this

Altered regulation of PDK4 expression promotes antiestrogen resistance in human breast cancer cells. / Walter, William; Thomalla, Jennifer; Bruhn, Josh; Fagan, Dedra H.; Zehowski, Cheryl; Yee, Douglas; Skildum, Andrew J.

In: SpringerPlus, Vol. 4, No. 1, 689, 01.12.2015, p. 1-13.

Research output: Contribution to journalArticle

Walter, William ; Thomalla, Jennifer ; Bruhn, Josh ; Fagan, Dedra H. ; Zehowski, Cheryl ; Yee, Douglas ; Skildum, Andrew J. / Altered regulation of PDK4 expression promotes antiestrogen resistance in human breast cancer cells. In: SpringerPlus. 2015 ; Vol. 4, No. 1. pp. 1-13.
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AB - Acquired or de novo resistance to the selective estrogen receptor modulators tamoxifen and fulvestrant (ICI) is a major barrier to successful treatment of breast cancer. Gene expression patterns in tamoxifen resistant (TamR-MCF-7) cells were compared to their parental cells (MCF-7L) to identify an aberrantly regulated metabolic pathway. TamR-MCF-7 cells are cross resistant to ICI and doxorubicin, and have increased mitochondrial DNA. A small subset of genes had altered expression in TamR-MCF-7 relative to MCF-7L cells. One of the genes, pyruvate dehydrogenase kinase-4 (PDK4), phosphorylates pyruvate dehydrogenase (PDH). PDK4 expression was elevated in TamR-MCF-7 cells; this result was also observed in a second model of acquired antiestrogen resistance. PDK4 expression is controlled in part by glucocorticoid response elements in the PDK4 gene promoter. In MCF-7L cells, PDK4 mRNA expression was insensitive to glucocorticoid receptor agonists, while dexamethasone dramatically increased PDK4 expression in TamR-MCF-7 cells. Using siRNA to knock down PDK4 expression increased TamR-MCF-7 sensitivity to ICI; in contrast adapting cells to growth in glucose depleted media did not affect ICI sensitivity. Despite TamR-MCF-7 cells high levels of PDK4 mRNA relative to MCF-7L, TamR-MCF-7 cells have increased PDH activity. Wild type MCF-7 cells are reported to be heterozygous for a G to A mutation that results in a substitution of threonine for alanine near PDK4′s catalytic site. We found loss of heterozygosity in TamR-MCF-7 cells; TamR-MCF-7 are homozygous for the wild type allele. These data support a role for altered regulation of PDH by PDK4 and altered substrate utilization in the development of drug resistance in human breast cancer cells.

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