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Nuclear factor I-C regulates E-cadherin via control of KLF4 in breast cancer.

Lee HK, Lee DS, Park JC - BMC Cancer (2015)

Bottom Line: We identified the downstream factors of NFI-C, such as KLF4 and E-cadherin, which play roles in EMT.NFI-C bound directly to the KLF4 promoter and stimulated KLF4 transcriptional activity, thereby regulating E-cadherin expression during tumorigenesis.The results indicate the important role of NFI-C in regulating KLF4 during tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Histology-Developmental Biology & Dental Research Institute, School of Dentistry, Seoul National University, 101 Daehagro, Chongro-gu, Seoul, 110-749, South Korea. h35071221@snu.ac.kr.

ABSTRACT

Background: Progression to metastasis is the leading cause of most cancer-related mortality; however, much remains to be understood about what facilitates the spread of tumor cells. In the present study, we describe a novel pathway in breast cancer that regulates epithelial-to-mesenchymal transition (EMT), motility, and invasiveness.

Methods: We examined nuclear factor I-C (NFI-C) expression in MCF10A human breast epithelial cells, MCF7 non-invasive breast cancer cells, and MDA-MB231 invasive breast cancer cells by real-time PCR and western blotting. To investigate the loss- and gain-function of NFI-C, we determined whether NFI-C regulated KLF4 expression by real-time PCR, western blotting, and promoter assay. To understand the biological functions of NFI-C, we observed cell invasion, migration, adhesion in human tumor cells by transwell assay, wound healing assay, quantitative RT-PCR, cell adhesion assay, western blotting, and immunohistochemistry.

Results: We identified the downstream factors of NFI-C, such as KLF4 and E-cadherin, which play roles in EMT. NFI-C is expressed in normal mammary gland or noninvasive breast cancer cells with epithelial characteristics. NFI-C overexpression induced expression of KLF4 and E-cadherin, but not Slug, in breast cancer cells. NFI-C bound directly to the KLF4 promoter and stimulated KLF4 transcriptional activity, thereby regulating E-cadherin expression during tumorigenesis. Cells overexpressing NFI-C maintained their epithelial differentiation status, which could drive mesenchymal-epithelial transition (MET) via the NFI-C-KLF4-E-cadherin axis in breast cancer cells. Consequently, NFI-C suppressed EMT, migration, and invasion in breast cancer cells.

Conclusions: Our study reveals a novel signaling pathway that is important during breast cancer tumorigenesis: the NFI-C-KLF4-E-cadherin pathway. The results indicate the important role of NFI-C in regulating KLF4 during tumorigenesis.

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Related in: MedlinePlus

TGF-β-mediated expression of NFI-C and E-cadherin. (A) Expression of NFI-C and E-cadherin in MCF10A human breast epithelial cells, MCF7 non-invasive breast cancer cells, and MDA-MB231 invasive breast cancer cells analyzed by western blotting. (B, C) Expression of NFI-C and E-cadherin in MCF10A cells treated with TGF-β and control MCF10A cells analyzed by real-time PCR (B) and western blotting (C).
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Fig1: TGF-β-mediated expression of NFI-C and E-cadherin. (A) Expression of NFI-C and E-cadherin in MCF10A human breast epithelial cells, MCF7 non-invasive breast cancer cells, and MDA-MB231 invasive breast cancer cells analyzed by western blotting. (B, C) Expression of NFI-C and E-cadherin in MCF10A cells treated with TGF-β and control MCF10A cells analyzed by real-time PCR (B) and western blotting (C).

Mentions: During tumor progression, carcinoma cells undergo EMT, a process in which polarized epithelial cells undergo dynamic changes to lose their epithelial characteristics and obtain a more motile fibroblastic phenotype, enabling them to proceed with invasion and metastasis [21]. To assess whether the presence of NFI-C in breast tumors has any physiologic relevance, we examined the NFI-C and E-cadherin protein expression in normal human breast epithelial cells, MCF10A, non-invasive breast cancer cells, MCF7, and invasive breast cancer cells, MDA-MB231 by western blotting. NFI-C expression correlated with cellular phenotype. Normal epithelial cells showed high expression of NFI-C and E-cadherin protein; whereas, the noninvasive and invasive cancer cells displayed little to no detectable expression of these two proteins (Figure 1A).Figure 1


Nuclear factor I-C regulates E-cadherin via control of KLF4 in breast cancer.

Lee HK, Lee DS, Park JC - BMC Cancer (2015)

TGF-β-mediated expression of NFI-C and E-cadherin. (A) Expression of NFI-C and E-cadherin in MCF10A human breast epithelial cells, MCF7 non-invasive breast cancer cells, and MDA-MB231 invasive breast cancer cells analyzed by western blotting. (B, C) Expression of NFI-C and E-cadherin in MCF10A cells treated with TGF-β and control MCF10A cells analyzed by real-time PCR (B) and western blotting (C).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359555&req=5

Fig1: TGF-β-mediated expression of NFI-C and E-cadherin. (A) Expression of NFI-C and E-cadherin in MCF10A human breast epithelial cells, MCF7 non-invasive breast cancer cells, and MDA-MB231 invasive breast cancer cells analyzed by western blotting. (B, C) Expression of NFI-C and E-cadherin in MCF10A cells treated with TGF-β and control MCF10A cells analyzed by real-time PCR (B) and western blotting (C).
Mentions: During tumor progression, carcinoma cells undergo EMT, a process in which polarized epithelial cells undergo dynamic changes to lose their epithelial characteristics and obtain a more motile fibroblastic phenotype, enabling them to proceed with invasion and metastasis [21]. To assess whether the presence of NFI-C in breast tumors has any physiologic relevance, we examined the NFI-C and E-cadherin protein expression in normal human breast epithelial cells, MCF10A, non-invasive breast cancer cells, MCF7, and invasive breast cancer cells, MDA-MB231 by western blotting. NFI-C expression correlated with cellular phenotype. Normal epithelial cells showed high expression of NFI-C and E-cadherin protein; whereas, the noninvasive and invasive cancer cells displayed little to no detectable expression of these two proteins (Figure 1A).Figure 1

Bottom Line: We identified the downstream factors of NFI-C, such as KLF4 and E-cadherin, which play roles in EMT.NFI-C bound directly to the KLF4 promoter and stimulated KLF4 transcriptional activity, thereby regulating E-cadherin expression during tumorigenesis.The results indicate the important role of NFI-C in regulating KLF4 during tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Histology-Developmental Biology & Dental Research Institute, School of Dentistry, Seoul National University, 101 Daehagro, Chongro-gu, Seoul, 110-749, South Korea. h35071221@snu.ac.kr.

ABSTRACT

Background: Progression to metastasis is the leading cause of most cancer-related mortality; however, much remains to be understood about what facilitates the spread of tumor cells. In the present study, we describe a novel pathway in breast cancer that regulates epithelial-to-mesenchymal transition (EMT), motility, and invasiveness.

Methods: We examined nuclear factor I-C (NFI-C) expression in MCF10A human breast epithelial cells, MCF7 non-invasive breast cancer cells, and MDA-MB231 invasive breast cancer cells by real-time PCR and western blotting. To investigate the loss- and gain-function of NFI-C, we determined whether NFI-C regulated KLF4 expression by real-time PCR, western blotting, and promoter assay. To understand the biological functions of NFI-C, we observed cell invasion, migration, adhesion in human tumor cells by transwell assay, wound healing assay, quantitative RT-PCR, cell adhesion assay, western blotting, and immunohistochemistry.

Results: We identified the downstream factors of NFI-C, such as KLF4 and E-cadherin, which play roles in EMT. NFI-C is expressed in normal mammary gland or noninvasive breast cancer cells with epithelial characteristics. NFI-C overexpression induced expression of KLF4 and E-cadherin, but not Slug, in breast cancer cells. NFI-C bound directly to the KLF4 promoter and stimulated KLF4 transcriptional activity, thereby regulating E-cadherin expression during tumorigenesis. Cells overexpressing NFI-C maintained their epithelial differentiation status, which could drive mesenchymal-epithelial transition (MET) via the NFI-C-KLF4-E-cadherin axis in breast cancer cells. Consequently, NFI-C suppressed EMT, migration, and invasion in breast cancer cells.

Conclusions: Our study reveals a novel signaling pathway that is important during breast cancer tumorigenesis: the NFI-C-KLF4-E-cadherin pathway. The results indicate the important role of NFI-C in regulating KLF4 during tumorigenesis.

Show MeSH
Related in: MedlinePlus