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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

Detection of growth factors involved in growth-promoting activity for MCF-7-E10 cells in BCTS. MCF-7-E10 cells were incubated with BCTS in the presence of anti-HGF antibody (a), AG1024, IGF-1R inhibitor (b), or AG1478, EGFR inhibitor (c), at the indicated concentrations. For anti-HGF antibody treatment, BCTS was pre-incubated with anti-HGF antibody for 30 min at room temperature and was then used for assay. Mouse IgG1 antibody was used as an isotype control. d The concentrations of HGF and EGF detected in BCTS were analyzed by immunoassay using Quantikine (R&D Systems, MN, USA). **, P < 0.01, ***, P < 0.001
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Fig5: Detection of growth factors involved in growth-promoting activity for MCF-7-E10 cells in BCTS. MCF-7-E10 cells were incubated with BCTS in the presence of anti-HGF antibody (a), AG1024, IGF-1R inhibitor (b), or AG1478, EGFR inhibitor (c), at the indicated concentrations. For anti-HGF antibody treatment, BCTS was pre-incubated with anti-HGF antibody for 30 min at room temperature and was then used for assay. Mouse IgG1 antibody was used as an isotype control. d The concentrations of HGF and EGF detected in BCTS were analyzed by immunoassay using Quantikine (R&D Systems, MN, USA). **, P < 0.01, ***, P < 0.001

Mentions: Growth-stimulating activity was heat labile and detectable in the fraction with an MW greater than 5 kDa (data not shown), suggesting that it could be derived from proteinous factors. Among various factors in the tumor microenvironment, HGF derived from stromal fibroblasts has been reported to stimulate growth of mouse mammary tumor cells in primary culture [27]; EGF and IGF-1 are known to activate ER via phosphorylation [18, 19]. To analyze the participation of these growth factors in tumor growth-stimulating activities found in BCTS, we first examined the effect of anti-HGF antibody on them. As shown in Fig. 5a, anti-HGF antibody, but not control IgG, effectively inhibited extract-stimulated growth of MCF-7-E10 cells. MCF-7 cells reportedly express c-Met, a receptor for HGF.Fig. 5


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)

Detection of growth factors involved in growth-promoting activity for MCF-7-E10 cells in BCTS. MCF-7-E10 cells were incubated with BCTS in the presence of anti-HGF antibody (a), AG1024, IGF-1R inhibitor (b), or AG1478, EGFR inhibitor (c), at the indicated concentrations. For anti-HGF antibody treatment, BCTS was pre-incubated with anti-HGF antibody for 30 min at room temperature and was then used for assay. Mouse IgG1 antibody was used as an isotype control. d The concentrations of HGF and EGF detected in BCTS were analyzed by immunoassay using Quantikine (R&D Systems, MN, USA). **, P < 0.01, ***, P < 0.001
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4150880&req=5

Fig5: Detection of growth factors involved in growth-promoting activity for MCF-7-E10 cells in BCTS. MCF-7-E10 cells were incubated with BCTS in the presence of anti-HGF antibody (a), AG1024, IGF-1R inhibitor (b), or AG1478, EGFR inhibitor (c), at the indicated concentrations. For anti-HGF antibody treatment, BCTS was pre-incubated with anti-HGF antibody for 30 min at room temperature and was then used for assay. Mouse IgG1 antibody was used as an isotype control. d The concentrations of HGF and EGF detected in BCTS were analyzed by immunoassay using Quantikine (R&D Systems, MN, USA). **, P < 0.01, ***, P < 0.001
Mentions: Growth-stimulating activity was heat labile and detectable in the fraction with an MW greater than 5 kDa (data not shown), suggesting that it could be derived from proteinous factors. Among various factors in the tumor microenvironment, HGF derived from stromal fibroblasts has been reported to stimulate growth of mouse mammary tumor cells in primary culture [27]; EGF and IGF-1 are known to activate ER via phosphorylation [18, 19]. To analyze the participation of these growth factors in tumor growth-stimulating activities found in BCTS, we first examined the effect of anti-HGF antibody on them. As shown in Fig. 5a, anti-HGF antibody, but not control IgG, effectively inhibited extract-stimulated growth of MCF-7-E10 cells. MCF-7 cells reportedly express c-Met, a receptor for HGF.Fig. 5

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