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FOXA1 expression affects the proliferation activity of luminal breast cancer stem cell populations.

Tachi K, Shiraishi A, Bando H, Yamashita T, Tsuboi I, Kato T, Hara H, Ohneda O - Cancer Sci. (2016)

Bottom Line: Using a quantitative PCR analysis, we found that mammosphere forming cells showed a higher expression of FOXA1 and stemness-related genes compared with adherent culture cells.As a result, we found no significant difference in the number of mammospheres but decreased colony formation in shFOXA1 MCF-7 cells compared with control.These results suggest that the expression of FOXA1 appears to be involved in the proliferation of immature BC cells rather than the induction of stemness-related genes and self-renewal potency of CSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast-Thyroid-Endocrine Surgery, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan.

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mRNA expression and Forkhead (FOXA1) levels compared between adherent culture samples and mammosphere culture samples. (a) mRNA expression levels of ER, FOXA1, SOX2, OCT4, and NANOG in each cell line are presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test. (b) Protein expression of FOXA1 in MCF‐7 (left) and BC#1 (right) cells compared between adherent culture and mammosphere culture by a Western blot analysis. (c) Relative expression of FOXA1 protein compared between cells grown in adherent culture and mammosphere culture. The expression levels were measured using Image J software and presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test.
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cas12870-fig-0003: mRNA expression and Forkhead (FOXA1) levels compared between adherent culture samples and mammosphere culture samples. (a) mRNA expression levels of ER, FOXA1, SOX2, OCT4, and NANOG in each cell line are presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test. (b) Protein expression of FOXA1 in MCF‐7 (left) and BC#1 (right) cells compared between adherent culture and mammosphere culture by a Western blot analysis. (c) Relative expression of FOXA1 protein compared between cells grown in adherent culture and mammosphere culture. The expression levels were measured using Image J software and presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test.

Mentions: First, the mRNA expression of ER, FOXA1, and stemness‐related genes NANOG, SOX2, and OCT4 were analyzed by quantitative PCR analyses.28, 40, 41, 42 There was no significant difference in the expression of ER between mammosphere and adhesion cultured cells. In contrast, the expression of FOXA1 under mammosphere culture was significantly higher than that under adherent culture (Fig. 3a). Similar to FOXA1 expression, stemness‐related genes were highly expressed in MCF‐7 and BC#1 cell lines under mammosphere culture compared to those under an adherent culture. Because lower mammosphere formation was observed in HCC1500 compared to that of MCF‐7 and BC#1 cell lines, we could not harvest sufficient samples for the analysis of gene expression. A Western blot analysis clearly indicated that FOXA1 protein expression was highly upregulated in both MCF‐7 and BC#1 cell lines under mammosphere culture compared to adherent culture (Fig. 3b).


FOXA1 expression affects the proliferation activity of luminal breast cancer stem cell populations.

Tachi K, Shiraishi A, Bando H, Yamashita T, Tsuboi I, Kato T, Hara H, Ohneda O - Cancer Sci. (2016)

mRNA expression and Forkhead (FOXA1) levels compared between adherent culture samples and mammosphere culture samples. (a) mRNA expression levels of ER, FOXA1, SOX2, OCT4, and NANOG in each cell line are presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test. (b) Protein expression of FOXA1 in MCF‐7 (left) and BC#1 (right) cells compared between adherent culture and mammosphere culture by a Western blot analysis. (c) Relative expression of FOXA1 protein compared between cells grown in adherent culture and mammosphere culture. The expression levels were measured using Image J software and presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test.
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cas12870-fig-0003: mRNA expression and Forkhead (FOXA1) levels compared between adherent culture samples and mammosphere culture samples. (a) mRNA expression levels of ER, FOXA1, SOX2, OCT4, and NANOG in each cell line are presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test. (b) Protein expression of FOXA1 in MCF‐7 (left) and BC#1 (right) cells compared between adherent culture and mammosphere culture by a Western blot analysis. (c) Relative expression of FOXA1 protein compared between cells grown in adherent culture and mammosphere culture. The expression levels were measured using Image J software and presented as the means ± SD from independent measurements. *P < 0.05, **P < 0.01, Student's t‐test.
Mentions: First, the mRNA expression of ER, FOXA1, and stemness‐related genes NANOG, SOX2, and OCT4 were analyzed by quantitative PCR analyses.28, 40, 41, 42 There was no significant difference in the expression of ER between mammosphere and adhesion cultured cells. In contrast, the expression of FOXA1 under mammosphere culture was significantly higher than that under adherent culture (Fig. 3a). Similar to FOXA1 expression, stemness‐related genes were highly expressed in MCF‐7 and BC#1 cell lines under mammosphere culture compared to those under an adherent culture. Because lower mammosphere formation was observed in HCC1500 compared to that of MCF‐7 and BC#1 cell lines, we could not harvest sufficient samples for the analysis of gene expression. A Western blot analysis clearly indicated that FOXA1 protein expression was highly upregulated in both MCF‐7 and BC#1 cell lines under mammosphere culture compared to adherent culture (Fig. 3b).

Bottom Line: Using a quantitative PCR analysis, we found that mammosphere forming cells showed a higher expression of FOXA1 and stemness-related genes compared with adherent culture cells.As a result, we found no significant difference in the number of mammospheres but decreased colony formation in shFOXA1 MCF-7 cells compared with control.These results suggest that the expression of FOXA1 appears to be involved in the proliferation of immature BC cells rather than the induction of stemness-related genes and self-renewal potency of CSCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast-Thyroid-Endocrine Surgery, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan.

Show MeSH
Related in: MedlinePlus