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GPR30-mediated estrogen signaling in Estrogen Receptor alpha and beta negative SKBR3 breast cancer cells

ABSTRACT: Gene expression analyses were carried out to identify genes regulated by 17-beta estradiol (E2) and Hydroxytamoxifen (OHT) through GPR30 in SKBR3 cells, a breast cancer cell-line which expresses GPR30 but lacks Estrogen Receptor alpha or beta. Keywords: Gene expression analysis, Non-genomic signaling in breast cancer cells. Gene expression analyses were done for control transfected SKBR3 cells: 1) Uninduced, 2) Induced with 10 microM OHT, 3) Induced with 1 microM E2 and 4) GPR30-antisense transfected cells induced with 10 microM OHT. The cells were induced for 1h and all the samples were collected in triplicates.

ORGANISM(S): Homo sapiens

SUBMITTER: Deo Prakash Pandey

PROVIDER: E-GEOD-11567 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Estrogenic GPR30 signalling induces proliferation and migration of breast cancer cells through CTGF.

Pandey Deo Prakash DP Lappano Rosamaria R Albanito Lidia L Madeo Antonio A Maggiolini Marcello M Picard Didier D

The EMBO journal 20090115 5

The steroid hormone oestrogen can signal through several receptors and pathways. Although the transcriptional responses mediated by the nuclear oestrogen receptors (ER) have been extensively characterized, the changes in gene expression elicited by signalling through the membrane-associated ER GPR30 have not been studied. We show here for ER-negative human breast cancer cells that the activation of GPR30 signalling by oestrogen or by hydroxytamoxifen (OHT), an ER antagonist but GPR30 agonist, in ...[more]

PMID: 19153601

Similar Datasets

GPR30-mediated estrogen signaling in Estrogen Receptor alpha and beta negative SKBR3 breast cancer cells

Project description:Gene expression analyses were carried out to identify genes regulated by 17-beta estradiol (E2) and Hydroxytamoxifen (OHT) through GPR30 in SKBR3 cells, a breast cancer cell-line which expresses GPR30 but lacks Estrogen Receptor alpha or beta. Keywords: Gene expression analysis, Non-genomic signaling in breast cancer cells.

2009-02-19 | GSE11567 | GEO

Effect of depletion of GPR30 on gene expression in female C57BL/6J mice

Project description:To test the cell-specific function of GPR30 in learning and memory, we generated global GPR30 knockout (KO) mice (Δ3102), astrocyte-specific GPR30 KO (AG-KO) mice and neuron-specific GPR30 KO (NG-KO) mice. We found that GPR30 in astrocytes but not neurons is required to maintain normal learning and memory. To further elucidate the signaling pathway by which GPR30 plays a role in astrocyte function, high-throughput RNA sequencing of hippocampal tissues from Δ3102, AG-KO, and NG-KO mice was performed.

2023-08-10 | GSE212893 | GEO

Time-course effect of estradiol and estradiol-BSA on early gene expression in SKBR3 cells

Project description:Estrogens have been reported to activate several processes via membrane binding to either classic estrogen receptors (ERs) or GPR30. We have used either estradiol or BSA-conjugated estradiol in order to initiate membrane-initiated actions and ICI 172,780 (ICI) or G15 to explore ER- and GPR30-related transcription. Our results show that the majority of G15-inhibited transcription is depending on ERs, as it is also inhibited by ICI. However, a small number of transcripts, related to specific actions/pathways is either exclusively inhibited by G15, providing evidence about a specific GPR30 signature, or not inhibited by ICI or G15 suggesting the existence of another, yet unidentified estrogen receptor. Cells after a 4h incubation with medium containing 10% charcoal stripped FBS were incubated with or without E2-BSA (10-6M) in the presence or absence of specific antagonists, in McCoys's 5A medium supplemented with 10% charcoal stripped FBS, for 3 hours. Total RNA was isolated using Nucleospin II columns (Macheray-Nagel, Dttren, Germany), according to the manufacturer’s instructions. RNA was labeled and hybridized according to the Affymetrix protocol (Affymetrix Gene-Chip Expression Analysis Technical Manual), using the HGU133A plus 2 chip, analyzing a total of 54675 transcripts. Signals were detected by an Affymetrix microarray chip reader.

2012-01-01 | E-GEOD-32669 | biostudies-arrayexpress

Time-course effect of estradiol and estradiol-BSA on early gene expression in MCF-7 cells

Project description:Estrogens have been reported to activate several processes via membrane binding to either classic estrogen receptors (ERs) or GPR30. We have used either estradiol or BSA-conjugated estradiol in order to initiate membrane-initiated actions and ICI 172,780 (ICI) or G15 to explore ER- and GPR30-related transcription. Our results show that the majority of G15-inhibited transcription is depending on ERs, as it is also inhibited by ICI. However, a small number of transcripts, related to specific actions/pathways is either exclusively inhibited by G15, providing evidence about a specific GPR30 signature, or not inhibited by ICI or G15 suggesting the existence of another, yet unidentified estrogen receptor. Cells after a 4h incubation with medium containing 10% charcoal stripped FBS were incubated with or without E2-BSA (10-6M) in the presence or absence of specific antagonists, in DMEM/F12 (1:1) supplemented with 10% charcoal stripped FBS, for 3 hours. Total RNA was isolated using Nucleospin II columns (Macheray-Nagel, Dttren, Germany), according to the manufacturer’s instructions. RNA was labeled and hybridized according to the Affymetrix protocol (Affymetrix Gene-Chip Expression Analysis Technical Manual), using the HGU133A plus 2 chip, analyzing a total of 54675 transcripts. Signals were detected by an Affymetrix microarray chip reader.

2012-01-01 | E-GEOD-32667 | biostudies-arrayexpress

Time-course effect of estradiol and estradiol-BSA on early gene expression in MDA-MB-231 cells

Project description:Estrogens have been reported to activate several processes via membrane binding to either classic estrogen receptors (ERs) or GPR30. We have used either estradiol or BSA-conjugated estradiol in order to initiate membrane-initiated actions and ICI 172,780 (ICI) or G15 to explore ER- and GPR30-related transcription. Our results show that the majority of G15-inhibited transcription is depending on ERs, as it is also inhibited by ICI. However, a small number of transcripts, related to specific actions/pathways is either exclusively inhibited by G15, providing evidence about a specific GPR30 signature, or not inhibited by ICI or G15 suggesting the existence of another, yet unidentified estrogen receptor. Cells after a 4h incubation with medium containing 10% charcoal stripped FBS were incubated with or without E2-BSA (10-6M) in the presence or absence of specific antagonists, in RPMI 1640 supplemented with 10% charcoal stripped FBS, for 3 hours. Total RNA was isolated using Nucleospin II columns (Macheray-Nagel, Dttren, Germany), according to the manufacturer’s instructions. RNA was labeled and hybridized according to the Affymetrix protocol (Affymetrix Gene-Chip Expression Analysis Technical Manual), using the HGU133A plus 2 chip, analyzing a total of 54675 transcripts. Signals were detected by an Affymetrix microarray chip reader.

2012-01-01 | E-GEOD-32668 | biostudies-arrayexpress

Time-course effect of estradiol and estradiol-BSA on early gene expression in T47D cells

Project description:Estrogens have been reported to activate several processes via membrane binding to either classic estrogen receptors (ERs) or GPR30. We have used either estradiol or BSA-conjugated estradiol in order to initiate membrane-initiated actions and ICI 172,780 (ICI) or G15 to explore ER- and GPR30-related transcription. Our results show that the majority of G15-inhibited transcription is depending on ERs, as it is also inhibited by ICI. However, a small number of transcripts, related to specific actions/pathways is either exclusively inhibited by G15, providing evidence about a specific GPR30 signature, or not inhibited by ICI or G15 suggesting the existence of another, yet unidentified estrogen receptor. Cells after a 4h incubation with medium containing 10% charcoal stripped FBS were incubated with or without E2-BSA (10-6M) in the presence or absence of specific antagonists, in RPMI 1640 supplemented with 10% charcoal stripped FBS, for 3 hours. Total RNA was isolated using Nucleospin II columns (Macheray-Nagel, Dttren, Germany), according to the manufacturer’s instructions. RNA was labeled and hybridized according to the Affymetrix protocol (Affymetrix Gene-Chip Expression Analysis Technical Manual), using the HGU133A plus 2 chip, analyzing a total of 54675 transcripts. Signals were detected by an Affymetrix microarray chip reader.

2012-01-01 | E-GEOD-32666 | biostudies-arrayexpress

RNAi expression experiments on Illumina BeadChip

Project description:Mouse embryonic stem cells (mES) were depleted of Klf2, Klf4, and Klf5 by ectopic expression of shRNA. Control RNAi was performed using shRNA against luciferase. At 2 days, 4 days and 6 days post-transfection of shRNA-encoding vectors, cells are harvested for RNA isolation. Keywords: RNAi expression, Mouse ES cells Knockdown: Klf2, Klf4, Klf5, Luciferase (Control);Time: 2 days post-transfection, 4 days post-transfection, 6 days post-transfection;Three independent experimental replicates for each experimental condition were performed.

2010-06-25 | E-GEOD-9775 | biostudies-arrayexpress

O-glycosylation inhibition and O-glycosylation mutants

Project description:This SuperSeries is composed of the following subset Series: GSE12180: Wild type 3 hours exposure to 1 microM OGT2468 GSE12181: Transcriptional profile of pmt1 pmt2 mutants GSE12182: Transcriptional profile of pmt1 pmt4 mutants GSE12193: Wild type 3 hours exposure to 0.1 microM OGT2468 Refer to individual Series

2010-09-29 | E-GEOD-12197 | biostudies-arrayexpress

Transcription profiling of human cells expressing FoxM1-ER fusion protein

Project description:Experiment description To identify FoxM1-regulated genes, we carried out gene expression profile analysis after inducible activation of a FoxM1-ER fusion protein using high-density human cDNA microarrays. We constructed a stable line of U2OS cells expressing full-length HA-FoxM1 fused at its C-terminus to the ligand-binding domain of the estrogen receptor (ER), mutated such that it specifically responds to 4-hydroxytamoxifen (4-OHT) but not to estrogen. In the absence of 4-OHT the activity of the ER fusion protein is turned off, but can be rapidly induced when cells are exposed to 4-OHT. We isolated total RNA from untreated cultures and cultures treated with 4-OHT for 6 or 16 hr. As activation of FoxM1 results in accumulation of cells in G2/M, we additionally analyzed the induction of gene expression by FoxM1 in G1/S-synchronized cells by a thymidine block. Total RNA derived from untreated or stimulated FoxM1-ER cells was amplified, reverse transcribed, labeled and hybridized to cDNA microarrays containing 18,000 cDNAs and expressed sequence tags (ESTs). Bound cDNA was detected using Cy3 or Cy5 dyes. In each independent experiment we included reverted color controls (Cy3 (4-OHT treated); Cy5 (untreated) or Cy3 (untreated); Cy5 (treated)) and the expression profile was expressed as a Cy5:Cy3 ratio.

2005-01-01 | E-MEXP-230 | biostudies-arrayexpress

Siomycin A treatment in prostate cancer cell lines PC3 and LNCaP

Project description:Siomycin A treatment in prostate cancer cell lines resulted in a concurrent inhibition of specific cell cycle genes and strongly impaired cell viability. Illumina microarray experiments were done using PC3 and LNCaP cell lines treated with 0.5 microM Siomaycin A either 24h or 48h in triplicates.

2012-10-19 | E-GEOD-36531 | biostudies-arrayexpress

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