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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

Correlations between growth-stimulating activity and clinicopathological characteristics, intrinsic subtypes and histological subtypes. 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 as described in Materials and Methods for triplicate experiments, and the growth-stimulating activities are shown as the ratios calculated relative to the control. Data are presented as mean ± SD of triplicate experiments. High growth-stimulating activity in specimens was associated with tumor size (a), intrinsic subtype (b), HER2 expression in ER-negative breast cancer (c) or histological classifications (d). Differences between groups were determined by two-sample t-test. P < 0.05 was considered statistically significant
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Fig4: Correlations between growth-stimulating activity and clinicopathological characteristics, intrinsic subtypes and histological subtypes. 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 as described in Materials and Methods for triplicate experiments, and the growth-stimulating activities are shown as the ratios calculated relative to the control. Data are presented as mean ± SD of triplicate experiments. High growth-stimulating activity in specimens was associated with tumor size (a), intrinsic subtype (b), HER2 expression in ER-negative breast cancer (c) or histological classifications (d). Differences between groups were determined by two-sample t-test. P < 0.05 was considered statistically significant

Mentions: We analyzed the relationships between ER-independent growth-stimulating activity detected in BCTS and clinicopathologic characteristics of the specimens’ donors (Fig. 4). Although BCTS growth-stimulating activity did not correlate with expression of ERα or PgR, stage, menopausal status, grade or nodal status (data not shown), specimens from tumors larger than 10 mm showed higher growth-stimulating activity than those smaller than 10 mm (Fig. 4a). Breast cancers are categorized into four intrinsic subtypes according to gene-expression profile: luminal A (ER + and/or PgR+, HER2–), luminal B (ER + and/or PgR+, HER2+), HER2 (ER–, PgR–, HER2+) and basal-type (ER–, PgR–, HER2–) [24, 25]. BCST derived from HER2 subtype showed slightly or significantly higher growth-stimulating activity than that from luminal B or basal types, respectively (Fig. 4b), suggesting that the tumor extracts of HER2 subtype have an abundance of growth factors stimulating their own receptors, including those of the ERBB family.Fig. 4


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)

Correlations between growth-stimulating activity and clinicopathological characteristics, intrinsic subtypes and histological subtypes. 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 as described in Materials and Methods for triplicate experiments, and the growth-stimulating activities are shown as the ratios calculated relative to the control. Data are presented as mean ± SD of triplicate experiments. High growth-stimulating activity in specimens was associated with tumor size (a), intrinsic subtype (b), HER2 expression in ER-negative breast cancer (c) or histological classifications (d). Differences between groups were determined by two-sample t-test. P < 0.05 was considered statistically significant
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4150880&req=5

Fig4: Correlations between growth-stimulating activity and clinicopathological characteristics, intrinsic subtypes and histological subtypes. 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 as described in Materials and Methods for triplicate experiments, and the growth-stimulating activities are shown as the ratios calculated relative to the control. Data are presented as mean ± SD of triplicate experiments. High growth-stimulating activity in specimens was associated with tumor size (a), intrinsic subtype (b), HER2 expression in ER-negative breast cancer (c) or histological classifications (d). Differences between groups were determined by two-sample t-test. P < 0.05 was considered statistically significant
Mentions: We analyzed the relationships between ER-independent growth-stimulating activity detected in BCTS and clinicopathologic characteristics of the specimens’ donors (Fig. 4). Although BCTS growth-stimulating activity did not correlate with expression of ERα or PgR, stage, menopausal status, grade or nodal status (data not shown), specimens from tumors larger than 10 mm showed higher growth-stimulating activity than those smaller than 10 mm (Fig. 4a). Breast cancers are categorized into four intrinsic subtypes according to gene-expression profile: luminal A (ER + and/or PgR+, HER2–), luminal B (ER + and/or PgR+, HER2+), HER2 (ER–, PgR–, HER2+) and basal-type (ER–, PgR–, HER2–) [24, 25]. BCST derived from HER2 subtype showed slightly or significantly higher growth-stimulating activity than that from luminal B or basal types, respectively (Fig. 4b), suggesting that the tumor extracts of HER2 subtype have an abundance of growth factors stimulating their own receptors, including those of the ERBB family.Fig. 4

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