Estrogen regulates vesicle trafficking gene expression in EFF-3, EFM-19 and MCF-7 breast cancer cells - PubMed (original) (raw)

Estrogen regulates vesicle trafficking gene expression in EFF-3, EFM-19 and MCF-7 breast cancer cells

Paul K Wright et al. Int J Clin Exp Pathol. 2009.

Abstract

Estrogens are critical mediators of breast tumorigenesis. This occurs via the action of estrogens on the estrogen receptor (ER), which regulates the transcriptome of breast cancer cells. Despite the long history of the search for estrogen-regulated genes in breast cancer, knowledge of the E2-regulated transcriptome and its effects is incomplete. We used Affymetrix GeneChips to profile the effects of estradiol on the expression of genes in EFF-3, EFM-19 and MCF-7 cells. In addition to many well-characterized estrogen-regulated genes, this identified a novel group of genes that have roles in vesicle trafficking, including exocytosis. Recent evidence in the literature supports a role for vesicle trafficking in tumorigenesis. We focused on five genes (SYTL5, RAB27B, SNX24, GALNT4 and SLC12A2/NKCC1/BSC2) and confirmed their estrogen-regulation using quantitative real-time PCR (qPCR). qPCR also demonstrated that these five genes were expressed in invasive breast carcinoma tissue. Immunohistochemistry showed expression of SYTL5 in cells of normal breast ductal epithelium, ductal carcinoma in-situ (DCIS) and invasive breast carcinoma. The results suggest that a significant effect of estrogens is to regulate the expression of genes that affect diverse aspects of vesicle trafficking including exocytosis.

Keywords: Breast cancer; RAB GTPase; estrogen; exocytosis; synaptotagmin-like protein; vesicle.

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Figures

Figure 1

Figure 1

qPCR demonstrating the mRNA expression of TFF1 in EFF-3, EFM-19, MCF-7, MDA-MB-231, Hs578T and SK-BR-3 cells after 48 hours E2-stimulation (E2) compared with steroid withdrawn (CONT) samples. (Error bars are standard deviation of three independent replicates.)

Figure 2

Figure 2

qPCR demonstrating E2-regulation of SYTL5, GALNT4, SNX24, RAB27B and SLC12A2. A. E2-regulation of all 5 genes in EFF-3, EFM-19 and MCF-7 cells showing mRNA fold change after 48 hours E2-treatment. B. The effects of E2-stimulation for 48 hours on SYTL5 mRNA expression on 6 breast cancer cell lines comparing steroid withdrawn (CONT) with E2-stimulated (E2) samples. C. SYTL5 mRNA expression in MCF-7 cells after E2-stimulation for 0 to 96 hours, comparing steroid withdrawn (CONT) with E2-stimulated (E2) samples. D. SYTL5 mRNA expression in MCF-7 cells after 48 hours in the presence of 0 – 10−7M E2. (Error bars are standard deviation of three independent replicates.)

Figure 3

Figure 3

mRNA expression of SYTL5, GALNT4, SNX24, RAB27B and SLC12A2 in invasive breast carcinoma tissue samples and MCF-7 cells. (• ERα+ tumors, ○ ERα- tumors, – mean mRNA expression, X MCF-7 cells. Y-axis: mRNA expression in relative units.)

Figure 4

Figure 4

SYTL5 protein expression in breast cancer. A. SYTL5 protein expression determined by Western blotting in MCF-7 and Hs578T cells. B–F. Immunohistochemistry showing SYTL5 protein expression in normal breast tissue (strong staining) (B), DCIS (strong staining) (C), ER- invasive breast carcinoma NST (moderate staining) (D) and ER+ invasive breast carcinoma NST (strong staining) (E and F). (Scale bar = 100μm)

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