Limits...
MicroRNA-148a is downregulated in gastric cancer, targets MMP7, and indicates tumor invasiveness and poor prognosis.

Sakamoto N, Naito Y, Oue N, Sentani K, Uraoka N, Zarni Oo H, Yanagihara K, Aoyagi K, Sasaki H, Yasui W - Cancer Sci. (2014)

Bottom Line: Downregulation of miR-148a was significantly correlated with an advanced clinical stage, lymph node metastasis, and poor clinical outcome.Custom oligonucleotide array analysis revealed that MMP7 expression was markedly downregulated in miR-148a-overexpressing GC cells; MMP7 was found to be a direct and functional target of miR-148a, participating in cell invasion.These results suggest that miR-148a contributes to the maintenance of homeostasis in normal stomach tissue and plays an important role in GC invasion by regulating MMP7 expression.

View Article: PubMed Central - PubMed

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

Show MeSH

Related in: MedlinePlus

MicroRNA-148a (miR-148a) expression and functional analysis. (a) Quantitative RT-PCR analysis of miR-148a in MKN-1 gastric cancer cells transfected with pre-miR-148a and HSC-57 cells transfected with anti-miR-148a. (b) Effect of miR-148a downregulation on cell invasion of MKN-1 cells. MKN-1 transfected with negative control miRNA and pre-miR-148a were incubated in Boyden chambers. (c) Effect of miR-148a downregulation on cell invasion of HSC-57 cells. HSC-57 transfected with negative control miRNA and anti-miR-148a were incubated in Boyden chambers. Results are mean ± SD of triplicate measurements. *P < 0.05; **P < 0.01; ***P < 0.001. N.S., not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4317816&req=5

fig02: MicroRNA-148a (miR-148a) expression and functional analysis. (a) Quantitative RT-PCR analysis of miR-148a in MKN-1 gastric cancer cells transfected with pre-miR-148a and HSC-57 cells transfected with anti-miR-148a. (b) Effect of miR-148a downregulation on cell invasion of MKN-1 cells. MKN-1 transfected with negative control miRNA and pre-miR-148a were incubated in Boyden chambers. (c) Effect of miR-148a downregulation on cell invasion of HSC-57 cells. HSC-57 transfected with negative control miRNA and anti-miR-148a were incubated in Boyden chambers. Results are mean ± SD of triplicate measurements. *P < 0.05; **P < 0.01; ***P < 0.001. N.S., not significant.

Mentions: To investigate the biological significance of miR-148a in GC, we carried out an MTT assay 4 days after altering miR-148a expression. In this experiment, we used MKN-1 cells transfected with miR-148a precursor and HSC-57 cells transfected with miR-148a inhibitor. MKN-1 and HSC-57 were selected as they possessed the lowest and highest endogenous miR-148a expression among nine GC cell lines, respectively. Successful overexpression and suppression of miR-148a in each treated cell line were confirmed by qRT-PCR (Fig. 2a). Cell growth of GC cells with deregulated miR-148a expression did not differ from that of cells transfected with control miRNA up to day 4 (data not shown). Next, a Transwell invasion assay was carried out. The invasiveness of miR-148a-overexpressing MKN-1 cells was reduced compared with the negative control miRNA-transfected MKN-1 cells (Fig. 2b). In contrast, the invasiveness of miR-148a-suppressed HSC-57 cells was greater than that of the negative control miRNA-transfected HSC-57 cells (Fig. 2c). These results are consistent with those of a previous report,(22) and indicate that miR-148a regulates some genes which in turn affect tumor invasion in GC cells.


MicroRNA-148a is downregulated in gastric cancer, targets MMP7, and indicates tumor invasiveness and poor prognosis.

Sakamoto N, Naito Y, Oue N, Sentani K, Uraoka N, Zarni Oo H, Yanagihara K, Aoyagi K, Sasaki H, Yasui W - Cancer Sci. (2014)

MicroRNA-148a (miR-148a) expression and functional analysis. (a) Quantitative RT-PCR analysis of miR-148a in MKN-1 gastric cancer cells transfected with pre-miR-148a and HSC-57 cells transfected with anti-miR-148a. (b) Effect of miR-148a downregulation on cell invasion of MKN-1 cells. MKN-1 transfected with negative control miRNA and pre-miR-148a were incubated in Boyden chambers. (c) Effect of miR-148a downregulation on cell invasion of HSC-57 cells. HSC-57 transfected with negative control miRNA and anti-miR-148a were incubated in Boyden chambers. Results are mean ± SD of triplicate measurements. *P < 0.05; **P < 0.01; ***P < 0.001. N.S., not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317816&req=5

fig02: MicroRNA-148a (miR-148a) expression and functional analysis. (a) Quantitative RT-PCR analysis of miR-148a in MKN-1 gastric cancer cells transfected with pre-miR-148a and HSC-57 cells transfected with anti-miR-148a. (b) Effect of miR-148a downregulation on cell invasion of MKN-1 cells. MKN-1 transfected with negative control miRNA and pre-miR-148a were incubated in Boyden chambers. (c) Effect of miR-148a downregulation on cell invasion of HSC-57 cells. HSC-57 transfected with negative control miRNA and anti-miR-148a were incubated in Boyden chambers. Results are mean ± SD of triplicate measurements. *P < 0.05; **P < 0.01; ***P < 0.001. N.S., not significant.
Mentions: To investigate the biological significance of miR-148a in GC, we carried out an MTT assay 4 days after altering miR-148a expression. In this experiment, we used MKN-1 cells transfected with miR-148a precursor and HSC-57 cells transfected with miR-148a inhibitor. MKN-1 and HSC-57 were selected as they possessed the lowest and highest endogenous miR-148a expression among nine GC cell lines, respectively. Successful overexpression and suppression of miR-148a in each treated cell line were confirmed by qRT-PCR (Fig. 2a). Cell growth of GC cells with deregulated miR-148a expression did not differ from that of cells transfected with control miRNA up to day 4 (data not shown). Next, a Transwell invasion assay was carried out. The invasiveness of miR-148a-overexpressing MKN-1 cells was reduced compared with the negative control miRNA-transfected MKN-1 cells (Fig. 2b). In contrast, the invasiveness of miR-148a-suppressed HSC-57 cells was greater than that of the negative control miRNA-transfected HSC-57 cells (Fig. 2c). These results are consistent with those of a previous report,(22) and indicate that miR-148a regulates some genes which in turn affect tumor invasion in GC cells.

Bottom Line: Downregulation of miR-148a was significantly correlated with an advanced clinical stage, lymph node metastasis, and poor clinical outcome.Custom oligonucleotide array analysis revealed that MMP7 expression was markedly downregulated in miR-148a-overexpressing GC cells; MMP7 was found to be a direct and functional target of miR-148a, participating in cell invasion.These results suggest that miR-148a contributes to the maintenance of homeostasis in normal stomach tissue and plays an important role in GC invasion by regulating MMP7 expression.

View Article: PubMed Central - PubMed

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

Show MeSH
Related in: MedlinePlus