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Detection of estrogen-independent growth-stimulating activity in breast cancer tissues: implication for tumor aggressiveness.

Yamaguchi Y, Seino Y, Takei H, Kurosumi M, Hayashi S - Cancer Microenviron (2013)

Bottom Line: In this study, we analyzed the effects of individual breast cancer extracts by our original system using an estrogen-signal reporter cell line, MCF-7-E10, which we previously established.High growth-promoting activities were seen in tumor regions of specimens with tumors > 10 mm in size, HER2 intrinsic subtype, and scirrhous and solid-tubular carcinoma histological subtypes.These data suggest that growth factors other than estrogen in the tumor extract significantly affect breast cancer aggressiveness in an estrogen-independent manner, and could be useful therapeutic targets.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Clinical Oncology, Saitama Cancer Center, 818 Komuro, Ina-machi, Saitama-ken, 362-0806, Japan, yamaguchi@cancer-c.pref.saitama.jp.

ABSTRACT
Estrogen and various growth factors affecting tumor behavior are present in the breast cancer microenvironment, but their comprehensive effects and signal crosstalks are different in each case. However, there is no system to evaluate the factors, detected in individual breast cancer cases, that regulate ER activity and tumor progression. In this study, we analyzed the effects of individual breast cancer extracts by our original system using an estrogen-signal reporter cell line, MCF-7-E10, which we previously established. MCF-7-E10 cell line is stably transfected by an estrogen response element (ERE)-green fluorescent protein (GFP) gene; it expresses GFP when estrogen receptors (ERs) are activated by estrogen or growth factor signal-mediated ER phosphorylation. Using this cell line, we analyzed the comprehensive effects of factors derived from breast cancer tissues on ER activity and growth of MCF-7-E10 cells for each case. We also analyzed relationships between these activities and clinicopathologic characteristics of patients who provided cancer specimens. The breast cancer extracts, which reflect the combined activities of growth factors present in individual cases, stimulated MCF-7-E10 cell growth in an estrogen-independent manner, and specifically stimulated growth of other breast cancer cell lines, regardless of ER expression. High growth-promoting activities were seen in tumor regions of specimens with tumors > 10 mm in size, HER2 intrinsic subtype, and scirrhous and solid-tubular carcinoma histological subtypes. Anti-human hepatocyte growth factor (HGF) antibody and an inhibitor for insulin-like growth factor-1 (IGF-1) receptor inhibited MCF-7-E10 cell growth by the breast cancer extracts, indicating that signal pathways via HGF or IGF-1 receptor significantly affect breast cancer. These data suggest that growth factors other than estrogen in the tumor extract significantly affect breast cancer aggressiveness in an estrogen-independent manner, and could be useful therapeutic targets.

No MeSH data available.


Related in: MedlinePlus

BCTS stimulated growth of MCF7-E10 cells in an estrogen-independent manner. a MCF7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate for 4 days. Cell growth was examined using a Cell Counting Kit-8 assay and ER activities are shown as the percentage of MCF-7-E10 cells expressing GFP. b, c MCF-7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate or the indicated concentrations in the presence or absence of anti-estrogen agents, fulvestrant (1 μM) or 4-hydroxy tamoxifen (4OHT, 1 μM), for 4 days. 17β-Estradiol (E2) was also tested at 1 nM or the indicated concentrations. Cell growth was examined using a Cell Counting Kit-8 assay. Data are presented as mean ± SD of triplicate experiments
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Fig3: BCTS stimulated growth of MCF7-E10 cells in an estrogen-independent manner. a MCF7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate for 4 days. Cell growth was examined using a Cell Counting Kit-8 assay and ER activities are shown as the percentage of MCF-7-E10 cells expressing GFP. b, c MCF-7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate or the indicated concentrations in the presence or absence of anti-estrogen agents, fulvestrant (1 μM) or 4-hydroxy tamoxifen (4OHT, 1 μM), for 4 days. 17β-Estradiol (E2) was also tested at 1 nM or the indicated concentrations. Cell growth was examined using a Cell Counting Kit-8 assay. Data are presented as mean ± SD of triplicate experiments

Mentions: To see if ER activation was required for BCTS-induced growth stimulation, we analyzed GFP expression in MCF-7-E10 cells, and found growth stimulation was not necessarily accompanied by ER activation (Fig. 3a). We next examined effects of anti-estrogen agents such as tamoxifen and fulvestrant on BCTS-induced growth stimulation, and found that high growth-stimulating activities were resistant to fulvestrant (Fig. 3b) and tamoxifen (Fig. 3c). These results indicate that, in addition to an ER-dependent pathway, BCTS stimulates breast cancer growth via an ER-independent pathway.Fig. 3


Detection of estrogen-independent growth-stimulating activity in breast cancer tissues: implication for tumor aggressiveness.

Yamaguchi Y, Seino Y, Takei H, Kurosumi M, Hayashi S - Cancer Microenviron (2013)

BCTS stimulated growth of MCF7-E10 cells in an estrogen-independent manner. a MCF7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate for 4 days. Cell growth was examined using a Cell Counting Kit-8 assay and ER activities are shown as the percentage of MCF-7-E10 cells expressing GFP. b, c MCF-7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate or the indicated concentrations in the presence or absence of anti-estrogen agents, fulvestrant (1 μM) or 4-hydroxy tamoxifen (4OHT, 1 μM), for 4 days. 17β-Estradiol (E2) was also tested at 1 nM or the indicated concentrations. Cell growth was examined using a Cell Counting Kit-8 assay. Data are presented as mean ± SD of triplicate experiments
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4150880&req=5

Fig3: BCTS stimulated growth of MCF7-E10 cells in an estrogen-independent manner. a MCF7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate for 4 days. Cell growth was examined using a Cell Counting Kit-8 assay and ER activities are shown as the percentage of MCF-7-E10 cells expressing GFP. b, c MCF-7-E10 cells were cultured with BCTS at 25 μg protein concentration in 150 μl medium per well in 96-well plate or the indicated concentrations in the presence or absence of anti-estrogen agents, fulvestrant (1 μM) or 4-hydroxy tamoxifen (4OHT, 1 μM), for 4 days. 17β-Estradiol (E2) was also tested at 1 nM or the indicated concentrations. Cell growth was examined using a Cell Counting Kit-8 assay. Data are presented as mean ± SD of triplicate experiments
Mentions: To see if ER activation was required for BCTS-induced growth stimulation, we analyzed GFP expression in MCF-7-E10 cells, and found growth stimulation was not necessarily accompanied by ER activation (Fig. 3a). We next examined effects of anti-estrogen agents such as tamoxifen and fulvestrant on BCTS-induced growth stimulation, and found that high growth-stimulating activities were resistant to fulvestrant (Fig. 3b) and tamoxifen (Fig. 3c). These results indicate that, in addition to an ER-dependent pathway, BCTS stimulates breast cancer growth via an ER-independent pathway.Fig. 3

Bottom Line: In this study, we analyzed the effects of individual breast cancer extracts by our original system using an estrogen-signal reporter cell line, MCF-7-E10, which we previously established.High growth-promoting activities were seen in tumor regions of specimens with tumors > 10 mm in size, HER2 intrinsic subtype, and scirrhous and solid-tubular carcinoma histological subtypes.These data suggest that growth factors other than estrogen in the tumor extract significantly affect breast cancer aggressiveness in an estrogen-independent manner, and could be useful therapeutic targets.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Clinical Oncology, Saitama Cancer Center, 818 Komuro, Ina-machi, Saitama-ken, 362-0806, Japan, yamaguchi@cancer-c.pref.saitama.jp.

ABSTRACT
Estrogen and various growth factors affecting tumor behavior are present in the breast cancer microenvironment, but their comprehensive effects and signal crosstalks are different in each case. However, there is no system to evaluate the factors, detected in individual breast cancer cases, that regulate ER activity and tumor progression. In this study, we analyzed the effects of individual breast cancer extracts by our original system using an estrogen-signal reporter cell line, MCF-7-E10, which we previously established. MCF-7-E10 cell line is stably transfected by an estrogen response element (ERE)-green fluorescent protein (GFP) gene; it expresses GFP when estrogen receptors (ERs) are activated by estrogen or growth factor signal-mediated ER phosphorylation. Using this cell line, we analyzed the comprehensive effects of factors derived from breast cancer tissues on ER activity and growth of MCF-7-E10 cells for each case. We also analyzed relationships between these activities and clinicopathologic characteristics of patients who provided cancer specimens. The breast cancer extracts, which reflect the combined activities of growth factors present in individual cases, stimulated MCF-7-E10 cell growth in an estrogen-independent manner, and specifically stimulated growth of other breast cancer cell lines, regardless of ER expression. High growth-promoting activities were seen in tumor regions of specimens with tumors > 10 mm in size, HER2 intrinsic subtype, and scirrhous and solid-tubular carcinoma histological subtypes. Anti-human hepatocyte growth factor (HGF) antibody and an inhibitor for insulin-like growth factor-1 (IGF-1) receptor inhibited MCF-7-E10 cell growth by the breast cancer extracts, indicating that signal pathways via HGF or IGF-1 receptor significantly affect breast cancer. These data suggest that growth factors other than estrogen in the tumor extract significantly affect breast cancer aggressiveness in an estrogen-independent manner, and could be useful therapeutic targets.

No MeSH data available.


Related in: MedlinePlus