Epigenetic-mediated upregulation of progesterone receptor B gene in endometrial cancer cell lines

Yuning Xiong, Sean Christopher Dowdy, Jesus Gonzalez Bosquet, Ying Zhao, Norman L. Eberhardt, Karl C. Podratz, Shi Wen Jiang

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Objectives. To determine if epigenetic interference can restore progesterone receptor-B (PR-B) expression in PR-B negative endometrial adenocarcinoma cell lines, and to characterize the kinetics of PR-B induction mediated by DNA methyltransferase and histone deacetylase inhibitors. Methods. The PR-B negative endometrioid cancer cell lines KLE and HEC-1B were used as study models. PR-B mRNA and protein expression levels were measured using real-time PCR and Western blot analysis, respectively. DNA methylation levels of the PR-B promoter were determined by methylation-specific PCR. Dose-response correlations and the duration of response to aza-deoxycytidine (ADC) and trichostatin A (TSA) were characterized. Cell responses to prolonged and repeated drug treatment were also examined. Results. Relatively low concentrations of ADC and TSA over a 24-h period induced PR-B expression. Furthermore, ADC and TSA acted synergistically to reactivate PR-B expression. Depending on the cell line used, PR-B mRNA was induced 10-110 fold. This elevated PR-B expression continued for 48 h after drug withdrawal. Sustained upregulation of PR-B mRNA and protein was observed during prolonged and repeated drug treatment. Conclusion. The epigenetically silenced PR-B gene remains sensitive to changes in DNA demethylation and histone acetylation in uterine adenocarcinoma cell lines. Treatment with ADC and/or TSA results in a robust and sustainable PR-B upregulation. These small molecule epigenetic modifying agents may be used to sensitize poorly differentiated, PR-B negative endometrial cancers to progestational therapy.

Original languageEnglish (US)
Pages (from-to)135-141
Number of pages7
JournalGynecologic Oncology
Volume99
Issue number1
DOIs
StatePublished - Oct 2005

Fingerprint

Endometrial Neoplasms
Epigenomics
Up-Regulation
Cell Line
trichostatin A
Genes
Deoxycytidine
progesterone receptor B
Messenger RNA
Adenocarcinoma
Pharmaceutical Preparations
Histone Deacetylase Inhibitors
DNA
Methyltransferases
DNA Methylation
Acetylation
Histones
Methylation
Real-Time Polymerase Chain Reaction
Proteins

Keywords

  • DNA methylation
  • Endometrial cancer
  • Epigenetics
  • Progesterone receptor

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Oncology

Cite this

Epigenetic-mediated upregulation of progesterone receptor B gene in endometrial cancer cell lines. / Xiong, Yuning; Dowdy, Sean Christopher; Bosquet, Jesus Gonzalez; Zhao, Ying; Eberhardt, Norman L.; Podratz, Karl C.; Jiang, Shi Wen.

In: Gynecologic Oncology, Vol. 99, No. 1, 10.2005, p. 135-141.

Research output: Contribution to journalArticle

Xiong, Yuning ; Dowdy, Sean Christopher ; Bosquet, Jesus Gonzalez ; Zhao, Ying ; Eberhardt, Norman L. ; Podratz, Karl C. ; Jiang, Shi Wen. / Epigenetic-mediated upregulation of progesterone receptor B gene in endometrial cancer cell lines. In: Gynecologic Oncology. 2005 ; Vol. 99, No. 1. pp. 135-141.
@article{18602d266daa49f0ab80f492f65d240e,
title = "Epigenetic-mediated upregulation of progesterone receptor B gene in endometrial cancer cell lines",
abstract = "Objectives. To determine if epigenetic interference can restore progesterone receptor-B (PR-B) expression in PR-B negative endometrial adenocarcinoma cell lines, and to characterize the kinetics of PR-B induction mediated by DNA methyltransferase and histone deacetylase inhibitors. Methods. The PR-B negative endometrioid cancer cell lines KLE and HEC-1B were used as study models. PR-B mRNA and protein expression levels were measured using real-time PCR and Western blot analysis, respectively. DNA methylation levels of the PR-B promoter were determined by methylation-specific PCR. Dose-response correlations and the duration of response to aza-deoxycytidine (ADC) and trichostatin A (TSA) were characterized. Cell responses to prolonged and repeated drug treatment were also examined. Results. Relatively low concentrations of ADC and TSA over a 24-h period induced PR-B expression. Furthermore, ADC and TSA acted synergistically to reactivate PR-B expression. Depending on the cell line used, PR-B mRNA was induced 10-110 fold. This elevated PR-B expression continued for 48 h after drug withdrawal. Sustained upregulation of PR-B mRNA and protein was observed during prolonged and repeated drug treatment. Conclusion. The epigenetically silenced PR-B gene remains sensitive to changes in DNA demethylation and histone acetylation in uterine adenocarcinoma cell lines. Treatment with ADC and/or TSA results in a robust and sustainable PR-B upregulation. These small molecule epigenetic modifying agents may be used to sensitize poorly differentiated, PR-B negative endometrial cancers to progestational therapy.",
keywords = "DNA methylation, Endometrial cancer, Epigenetics, Progesterone receptor",
author = "Yuning Xiong and Dowdy, {Sean Christopher} and Bosquet, {Jesus Gonzalez} and Ying Zhao and Eberhardt, {Norman L.} and Podratz, {Karl C.} and Jiang, {Shi Wen}",
year = "2005",
month = "10",
doi = "10.1016/j.ygyno.2005.05.035",
language = "English (US)",
volume = "99",
pages = "135--141",
journal = "Gynecologic Oncology",
issn = "0090-8258",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Epigenetic-mediated upregulation of progesterone receptor B gene in endometrial cancer cell lines

AU - Xiong, Yuning

AU - Dowdy, Sean Christopher

AU - Bosquet, Jesus Gonzalez

AU - Zhao, Ying

AU - Eberhardt, Norman L.

AU - Podratz, Karl C.

AU - Jiang, Shi Wen

PY - 2005/10

Y1 - 2005/10

N2 - Objectives. To determine if epigenetic interference can restore progesterone receptor-B (PR-B) expression in PR-B negative endometrial adenocarcinoma cell lines, and to characterize the kinetics of PR-B induction mediated by DNA methyltransferase and histone deacetylase inhibitors. Methods. The PR-B negative endometrioid cancer cell lines KLE and HEC-1B were used as study models. PR-B mRNA and protein expression levels were measured using real-time PCR and Western blot analysis, respectively. DNA methylation levels of the PR-B promoter were determined by methylation-specific PCR. Dose-response correlations and the duration of response to aza-deoxycytidine (ADC) and trichostatin A (TSA) were characterized. Cell responses to prolonged and repeated drug treatment were also examined. Results. Relatively low concentrations of ADC and TSA over a 24-h period induced PR-B expression. Furthermore, ADC and TSA acted synergistically to reactivate PR-B expression. Depending on the cell line used, PR-B mRNA was induced 10-110 fold. This elevated PR-B expression continued for 48 h after drug withdrawal. Sustained upregulation of PR-B mRNA and protein was observed during prolonged and repeated drug treatment. Conclusion. The epigenetically silenced PR-B gene remains sensitive to changes in DNA demethylation and histone acetylation in uterine adenocarcinoma cell lines. Treatment with ADC and/or TSA results in a robust and sustainable PR-B upregulation. These small molecule epigenetic modifying agents may be used to sensitize poorly differentiated, PR-B negative endometrial cancers to progestational therapy.

AB - Objectives. To determine if epigenetic interference can restore progesterone receptor-B (PR-B) expression in PR-B negative endometrial adenocarcinoma cell lines, and to characterize the kinetics of PR-B induction mediated by DNA methyltransferase and histone deacetylase inhibitors. Methods. The PR-B negative endometrioid cancer cell lines KLE and HEC-1B were used as study models. PR-B mRNA and protein expression levels were measured using real-time PCR and Western blot analysis, respectively. DNA methylation levels of the PR-B promoter were determined by methylation-specific PCR. Dose-response correlations and the duration of response to aza-deoxycytidine (ADC) and trichostatin A (TSA) were characterized. Cell responses to prolonged and repeated drug treatment were also examined. Results. Relatively low concentrations of ADC and TSA over a 24-h period induced PR-B expression. Furthermore, ADC and TSA acted synergistically to reactivate PR-B expression. Depending on the cell line used, PR-B mRNA was induced 10-110 fold. This elevated PR-B expression continued for 48 h after drug withdrawal. Sustained upregulation of PR-B mRNA and protein was observed during prolonged and repeated drug treatment. Conclusion. The epigenetically silenced PR-B gene remains sensitive to changes in DNA demethylation and histone acetylation in uterine adenocarcinoma cell lines. Treatment with ADC and/or TSA results in a robust and sustainable PR-B upregulation. These small molecule epigenetic modifying agents may be used to sensitize poorly differentiated, PR-B negative endometrial cancers to progestational therapy.

KW - DNA methylation

KW - Endometrial cancer

KW - Epigenetics

KW - Progesterone receptor

UR - http://www.scopus.com/inward/record.url?scp=25144454885&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=25144454885&partnerID=8YFLogxK

U2 - 10.1016/j.ygyno.2005.05.035

DO - 10.1016/j.ygyno.2005.05.035

M3 - Article

C2 - 16024066

AN - SCOPUS:25144454885

VL - 99

SP - 135

EP - 141

JO - Gynecologic Oncology

JF - Gynecologic Oncology

SN - 0090-8258

IS - 1

ER -