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MicroRNA-143 regulates collagen type III expression in stromal fibroblasts of scirrhous type gastric cancer.

Naito Y, Sakamoto N, Oue N, Yashiro M, Sentani K, Yanagihara K, Hirakawa K, Yasui W - Cancer Sci. (2014)

Bottom Line: In stromal cells, miR-143 enhanced collagen type III expression in normal gastric fibroblasts and cancer-associated fibroblasts through activation of transforming growth factor-β)/SMAD signaling.Furthermore, high miR-143 expression in GC was associated with worse cancer-specific mortality (P = 0.0141).These data suggest that miR-143 regulates fibrosis of scirrhous type GC through induction of collagen expression in stromal fibroblasts and that miR-143 expression serves as a prognostic marker of GC.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

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Regulation of collagen type III expression by microRNA-143 (miR-143). (a) Collagen type III expression was assessed by immunohistochemical analysis of scirrhous type gastric cancer (GC) tissue. (b) Collagen type III mRNA expression levels were evaluated in GC and fibroblasts. Normal gastric fibroblasts and cancer-associated fibroblasts (NF-38 and CaF-38, respectively) were transfected with negative control miRNA or precursor miR-143 or miR-143 inhibitor, and (c) quantitative RT-PCR, (d) Western blot, and (e) cell staining were carried out for collagen type III expression. Scale bars: 50 μm. (f) Proliferation activity after coculture of HSC-44PE GC cells and CaF-38 with miR-143 inhibitor or negative control. Proliferation activity of HSC-44PE was assessed by percentages of BrdU/CAM5.2-positive cells. (g) Proliferation activity after coculture of HSC-44PE and CaF-38 with COL3A1 siRNA or negative control. Collagen type III expression level was determined by Western blot analysis. Results are mean ± SE of triplicate measurements. *P < 0.05.
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fig03: Regulation of collagen type III expression by microRNA-143 (miR-143). (a) Collagen type III expression was assessed by immunohistochemical analysis of scirrhous type gastric cancer (GC) tissue. (b) Collagen type III mRNA expression levels were evaluated in GC and fibroblasts. Normal gastric fibroblasts and cancer-associated fibroblasts (NF-38 and CaF-38, respectively) were transfected with negative control miRNA or precursor miR-143 or miR-143 inhibitor, and (c) quantitative RT-PCR, (d) Western blot, and (e) cell staining were carried out for collagen type III expression. Scale bars: 50 μm. (f) Proliferation activity after coculture of HSC-44PE GC cells and CaF-38 with miR-143 inhibitor or negative control. Proliferation activity of HSC-44PE was assessed by percentages of BrdU/CAM5.2-positive cells. (g) Proliferation activity after coculture of HSC-44PE and CaF-38 with COL3A1 siRNA or negative control. Collagen type III expression level was determined by Western blot analysis. Results are mean ± SE of triplicate measurements. *P < 0.05.

Mentions: Because miR-143 was found to be expressed in stromal fibroblasts but not in cancer cells, we sought to investigate the function of miR-143 in stromal fibroblasts. Scirrhous type GC produces abundant collagen and thus promotes fibrosis.(31,32) We previously reported that collagen type III expression is associated with scirrhous type GC.(6,7) We first examined collagen type III expression in scirrhous type GC tissue by immunostaining and, as expected, collagen type III was detected in fibrillar bundles of scirrhous type GC (Fig. 3a). Collagen type III mRNA expression was examined in GC and fibroblasts by qRT-PCR. High levels of collagen type III mRNA expression were observed in stromal fibroblasts that retained high miR-143 expression (Fig. 3b). To assess the relation between collagen type III and miR-143, NF-38 and CaF-38 were selected because they had the highest miR-143 and collagen type III mRNA expression (Fig. 3b). Transfection of miR-143 inhibitor significantly suppressed collagen type III expression (Fig. 3c–e). In contrast, transfection of miR-143 precursor sustained or increased collagen type III expression (Fig. 3c–e). These data suggest that miR-143 positively regulates collagen type III expression in stromal fibroblasts of scirrhous type GC.


MicroRNA-143 regulates collagen type III expression in stromal fibroblasts of scirrhous type gastric cancer.

Naito Y, Sakamoto N, Oue N, Yashiro M, Sentani K, Yanagihara K, Hirakawa K, Yasui W - Cancer Sci. (2014)

Regulation of collagen type III expression by microRNA-143 (miR-143). (a) Collagen type III expression was assessed by immunohistochemical analysis of scirrhous type gastric cancer (GC) tissue. (b) Collagen type III mRNA expression levels were evaluated in GC and fibroblasts. Normal gastric fibroblasts and cancer-associated fibroblasts (NF-38 and CaF-38, respectively) were transfected with negative control miRNA or precursor miR-143 or miR-143 inhibitor, and (c) quantitative RT-PCR, (d) Western blot, and (e) cell staining were carried out for collagen type III expression. Scale bars: 50 μm. (f) Proliferation activity after coculture of HSC-44PE GC cells and CaF-38 with miR-143 inhibitor or negative control. Proliferation activity of HSC-44PE was assessed by percentages of BrdU/CAM5.2-positive cells. (g) Proliferation activity after coculture of HSC-44PE and CaF-38 with COL3A1 siRNA or negative control. Collagen type III expression level was determined by Western blot analysis. Results are mean ± SE of triplicate measurements. *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig03: Regulation of collagen type III expression by microRNA-143 (miR-143). (a) Collagen type III expression was assessed by immunohistochemical analysis of scirrhous type gastric cancer (GC) tissue. (b) Collagen type III mRNA expression levels were evaluated in GC and fibroblasts. Normal gastric fibroblasts and cancer-associated fibroblasts (NF-38 and CaF-38, respectively) were transfected with negative control miRNA or precursor miR-143 or miR-143 inhibitor, and (c) quantitative RT-PCR, (d) Western blot, and (e) cell staining were carried out for collagen type III expression. Scale bars: 50 μm. (f) Proliferation activity after coculture of HSC-44PE GC cells and CaF-38 with miR-143 inhibitor or negative control. Proliferation activity of HSC-44PE was assessed by percentages of BrdU/CAM5.2-positive cells. (g) Proliferation activity after coculture of HSC-44PE and CaF-38 with COL3A1 siRNA or negative control. Collagen type III expression level was determined by Western blot analysis. Results are mean ± SE of triplicate measurements. *P < 0.05.
Mentions: Because miR-143 was found to be expressed in stromal fibroblasts but not in cancer cells, we sought to investigate the function of miR-143 in stromal fibroblasts. Scirrhous type GC produces abundant collagen and thus promotes fibrosis.(31,32) We previously reported that collagen type III expression is associated with scirrhous type GC.(6,7) We first examined collagen type III expression in scirrhous type GC tissue by immunostaining and, as expected, collagen type III was detected in fibrillar bundles of scirrhous type GC (Fig. 3a). Collagen type III mRNA expression was examined in GC and fibroblasts by qRT-PCR. High levels of collagen type III mRNA expression were observed in stromal fibroblasts that retained high miR-143 expression (Fig. 3b). To assess the relation between collagen type III and miR-143, NF-38 and CaF-38 were selected because they had the highest miR-143 and collagen type III mRNA expression (Fig. 3b). Transfection of miR-143 inhibitor significantly suppressed collagen type III expression (Fig. 3c–e). In contrast, transfection of miR-143 precursor sustained or increased collagen type III expression (Fig. 3c–e). These data suggest that miR-143 positively regulates collagen type III expression in stromal fibroblasts of scirrhous type GC.

Bottom Line: In stromal cells, miR-143 enhanced collagen type III expression in normal gastric fibroblasts and cancer-associated fibroblasts through activation of transforming growth factor-β)/SMAD signaling.Furthermore, high miR-143 expression in GC was associated with worse cancer-specific mortality (P = 0.0141).These data suggest that miR-143 regulates fibrosis of scirrhous type GC through induction of collagen expression in stromal fibroblasts and that miR-143 expression serves as a prognostic marker of GC.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

Show MeSH
Related in: MedlinePlus