Dataset Information

RHBDD2 silencing and gene expression profiling of breast cancer cells

ABSTRACT: In previous studies, we identified a distantly related rhomboid homologue gene known as RHBDD2 (Rhomboid domain containing 2) to be markedly overexpressed in the advanced stages of the breast and colorectal cancer diseases. In order to identify RHBDD2 modulated pathways, we analyzed two breast cancer cell lines (MCF7 and T47D) from control and RHBDD2-siRNA transient gene silencing followed by gene expression profiling analysis using the whole genome Toray 3D-GeneTM Human Oligo Chip. Statistical analysis of the Toray's 3D gene expression profiling data identified 566 commonly differentially expressed genes in association to the RHBDD2 knockdown in both breast cancer cell lines. Among the statistically significant over-represented biological process, we found the apoptosis, cell cycle and response to DNA damage process related genes. In addition, categories of genes found in the ubiquitin-proteasome and oxidative phosphorylation were also highly enriched related genes in the commonly deregulated gene list. We further used a lentivirus-based system (shRNA-pLKO.1) for stable silencing of RHBDD2 mRNA in the T47D breast cancer cell line. Using a staurosporine-induced apoptosis model, we demonstrate that RHBDD2 abrogation resulted in an apoptosis-resistant phenotype of T47D breast cancer cell line. These data are in line with a recent study, suggesting that RHBDD2 expression could be up-modulated in response to 5FU-induced apoptosis in colorectal cancer cells. Taken together, these data suggest that RHBDD2 could be involved in the modulation of the programmed cell death in cancer cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE43015 | GEO | 2013/04/10

SECONDARY ACCESSION(S): PRJNA184057

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:17Î²-HSD1 expression modulates T47D transcript profile in in steroid-deprived medium. The enzyme specifically regulates apoptosis and cancer-related genes in T47D cells cultured in steroid-deprived medium. Genes that primarily involved in Cell cycle progression, such as the Cyclin A2 gene, CCNA2, are generally down-regulated whereas most genes involved in apoptosis and cell death, such as the proapoptotic gene XAF1 and FGF12, are on the contrary up-regulated by 17Î²-HSD1 knockdown, and 21% of the modulated genes belong to this latter functional category. This correborates with its role previously shown in increasing breast cancer cell proliferation (Aka et al, 2010) and indicates that in addition to its direct role as cell proliferation via incresing E2 and decreasing DHT, 17Î²-HSD1 may also be involved in oncogenesis by favoring its anti-apoptosis pathway in breast cancer cells 17Î²-HSD1 may also be involved in oncogenesis by favoring anti-apoptosis pathway and/or inhibiting cell survival pathway. We tested the ability of estrogen to induce or repress endogenous genes of T47D by microarray analysis. A total of 131 genes were found to increase or decrease 1.5-fold or higher in response to 17 beta-estradiol (1 nM for 48 h). Besides, the gene regulation occuring in steroid-deprived conditions showed that 17Î²-HSD1 modulates gene expression in steroid-independent manners. Our study thus demonstrates that 17Î²-HSD1 modulates the E2-independent transcription of endogenous genes in T47D cells. The E2-dependent transcription as well as the E2-independent transcription are significantly modulated by 17Î²-HSD1 in breast cancer cells We first report here that breast cancer cell transcript profile is independtly modulated by both 17Î²-HSD1 and its principale product estradiol. Microarrays was used to study the effect of 17Î²-HSD1 gene knockdown on gene expression and on endogenous ERG in T47D cells. The sense and antisense sequences of three 17Î²-HSD1 siRNAs were selected and synthesized. Scramble siRNA was used as control siRNA. Two days before transfection, T47D cells in T75 were cultured in steroid deprived medium. On the transfection day, cells in T75 flasks were trypsined and ressuspended in a fresh charcoal-treated medium. Cells were then reverse-transfected with 17Î²-HSD1 specific siRNAs or with negative control siRNA.Two days (48 hours) after transfection, cell culture media were replaced by fresh charcoal-treated medium containing either the steroid E2 (1 nM) or ethanol as a vehicle control, and cells were incubated for two more days before RNA extraction.

Dataset Information

RHBDD2 silencing and gene expression profiling of breast cancer cells

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets