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miR-203 Inhibits Frizzled-2 Expression via CD82/KAI1 Expression in Human Lung Carcinoma Cells.

Mine M, Yamaguchi K, Sugiura T, Chigita S, Yoshihama N, Yoshihama R, Hiyake N, Kobayashi Y, Mori Y - PLoS ONE (2015)

Bottom Line: In this study, we investigated the mechanism through which CD82 inhibited FZD expression by examining the effects of microRNAs (miRNAs).Among these miRNAs, CD82 caused upregulation of miR-203 (by 2.095-fold) and downregulation of miR-338-3p (by 0.354-fold) as compared with control cells.Therefore, these findings suggested that CD82 enhanced the expression of miR-203 and directly downregulate FZD2 expression, suppressing cancer metastasis by inhibition of the Wnt signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Division of Maxillofacial Diagnostic and Surgical Sciences, Department of Oral and Maxillofacial Surgery, Graduate School of Dental Science, Kyushu University Maidashi, Higashi-ku, Fukuoka, Japan.

ABSTRACT
CD82/KAI1, a member of the tetraspanin superfamily, is a suppressor of metastasis and CD82 inhibits canonical Wnt signaling via downregulation of several Frizzled (FZD) isoforms, resulting in accumulation of β-catenin at the cell membrane. In this study, we investigated the mechanism through which CD82 inhibited FZD expression by examining the effects of microRNAs (miRNAs). The miRanda algorithm predicted 11 miRNAs from FZD sequences. Among these miRNAs, CD82 caused upregulation of miR-203 (by 2.095-fold) and downregulation of miR-338-3p (by 0.354-fold) as compared with control cells. Transfection with miR-203 and miR338-3p mimics or inhibitors revealed that miR-203 downregulated FZD2 mRNA (by 0.268-fold) and protein expression (by 0.701-fold). Moreover, transfection with the miR-203 mimic also inhibited cell migration. Therefore, these findings suggested that CD82 enhanced the expression of miR-203 and directly downregulate FZD2 expression, suppressing cancer metastasis by inhibition of the Wnt signaling pathway.

No MeSH data available.


Related in: MedlinePlus

Transfection with miRNA mimics and hairpin inhibitors.Total RNA was isolated from h1299 cells at 48 h after transfection with miRNA mimics or miRNA hairpin inhibitors, and miRNA levels were then analyzed by real-time RT-PCR. RNU6B was used as an internal reference gene. Experiments were performed in triplicate, and relative miRNA levels (zeo/control inhibitor = 1) were averaged. The asterisks in the figure indicate statistically significant differences (*p < 0.01) between the two values. Data are presented as the means ± SDs.
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pone.0131350.g002: Transfection with miRNA mimics and hairpin inhibitors.Total RNA was isolated from h1299 cells at 48 h after transfection with miRNA mimics or miRNA hairpin inhibitors, and miRNA levels were then analyzed by real-time RT-PCR. RNU6B was used as an internal reference gene. Experiments were performed in triplicate, and relative miRNA levels (zeo/control inhibitor = 1) were averaged. The asterisks in the figure indicate statistically significant differences (*p < 0.01) between the two values. Data are presented as the means ± SDs.

Mentions: To examine the functional effects of miR-338-3p and miR-203 on the regulation of FZDs, we transiently transfected h1299/zeo and h1299/CD82 cells with miR-338-3p or miR-203 mimics or inhibitors. Forty-eight hours after transfection, miRNA levels of miR-338-3p and miR-203 were significantly decreased in h1299/zeo cells transfected with the miR-338-3p inhibitor and h1299/CD82 cells transfected with the miR-203 inhibitor (Fig 2A and 2D). Transfection with miR-338-3p and miR-203 mimics significantly increased the miRNA levels in h1299/CD82 cells transfected with miR-338-3p mimic and in h1299/zeo cells transfected with the miR-203 mimic (Fig 2B and 2C).


miR-203 Inhibits Frizzled-2 Expression via CD82/KAI1 Expression in Human Lung Carcinoma Cells.

Mine M, Yamaguchi K, Sugiura T, Chigita S, Yoshihama N, Yoshihama R, Hiyake N, Kobayashi Y, Mori Y - PLoS ONE (2015)

Transfection with miRNA mimics and hairpin inhibitors.Total RNA was isolated from h1299 cells at 48 h after transfection with miRNA mimics or miRNA hairpin inhibitors, and miRNA levels were then analyzed by real-time RT-PCR. RNU6B was used as an internal reference gene. Experiments were performed in triplicate, and relative miRNA levels (zeo/control inhibitor = 1) were averaged. The asterisks in the figure indicate statistically significant differences (*p < 0.01) between the two values. Data are presented as the means ± SDs.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4488846&req=5

pone.0131350.g002: Transfection with miRNA mimics and hairpin inhibitors.Total RNA was isolated from h1299 cells at 48 h after transfection with miRNA mimics or miRNA hairpin inhibitors, and miRNA levels were then analyzed by real-time RT-PCR. RNU6B was used as an internal reference gene. Experiments were performed in triplicate, and relative miRNA levels (zeo/control inhibitor = 1) were averaged. The asterisks in the figure indicate statistically significant differences (*p < 0.01) between the two values. Data are presented as the means ± SDs.
Mentions: To examine the functional effects of miR-338-3p and miR-203 on the regulation of FZDs, we transiently transfected h1299/zeo and h1299/CD82 cells with miR-338-3p or miR-203 mimics or inhibitors. Forty-eight hours after transfection, miRNA levels of miR-338-3p and miR-203 were significantly decreased in h1299/zeo cells transfected with the miR-338-3p inhibitor and h1299/CD82 cells transfected with the miR-203 inhibitor (Fig 2A and 2D). Transfection with miR-338-3p and miR-203 mimics significantly increased the miRNA levels in h1299/CD82 cells transfected with miR-338-3p mimic and in h1299/zeo cells transfected with the miR-203 mimic (Fig 2B and 2C).

Bottom Line: In this study, we investigated the mechanism through which CD82 inhibited FZD expression by examining the effects of microRNAs (miRNAs).Among these miRNAs, CD82 caused upregulation of miR-203 (by 2.095-fold) and downregulation of miR-338-3p (by 0.354-fold) as compared with control cells.Therefore, these findings suggested that CD82 enhanced the expression of miR-203 and directly downregulate FZD2 expression, suppressing cancer metastasis by inhibition of the Wnt signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Division of Maxillofacial Diagnostic and Surgical Sciences, Department of Oral and Maxillofacial Surgery, Graduate School of Dental Science, Kyushu University Maidashi, Higashi-ku, Fukuoka, Japan.

ABSTRACT
CD82/KAI1, a member of the tetraspanin superfamily, is a suppressor of metastasis and CD82 inhibits canonical Wnt signaling via downregulation of several Frizzled (FZD) isoforms, resulting in accumulation of β-catenin at the cell membrane. In this study, we investigated the mechanism through which CD82 inhibited FZD expression by examining the effects of microRNAs (miRNAs). The miRanda algorithm predicted 11 miRNAs from FZD sequences. Among these miRNAs, CD82 caused upregulation of miR-203 (by 2.095-fold) and downregulation of miR-338-3p (by 0.354-fold) as compared with control cells. Transfection with miR-203 and miR338-3p mimics or inhibitors revealed that miR-203 downregulated FZD2 mRNA (by 0.268-fold) and protein expression (by 0.701-fold). Moreover, transfection with the miR-203 mimic also inhibited cell migration. Therefore, these findings suggested that CD82 enhanced the expression of miR-203 and directly downregulate FZD2 expression, suppressing cancer metastasis by inhibition of the Wnt signaling pathway.

No MeSH data available.


Related in: MedlinePlus