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Downregulation of miR-106b induced breast cancer cell invasion and motility in association with overexpression of matrix metalloproteinase 2.

Ni X, Xia T, Zhao Y, Zhou W, Wu N, Liu X, Ding Q, Zha X, Sha J, Wang S - Cancer Sci. (2013)

Bottom Line: MMP2 was shown to be a direct target of miR-106b.Both gain- and loss-of-function studies showed that MMP2 could promote the migration and invasion of BC cells and that miR-106b could suppress both.The blockage of MMP2 by RNA interference mimicked the anti-migration and anti-invasion effects of miR-106b, and introduction of MMP2 antagonized the function of miR-106b.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China; State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, China.

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Matrix metalloproteinase 2 (MMP2) is the direct target of miR-106b. (a) Real-time polymerase chain reaction (PCR) analysis demonstrated higher levels of miR-106b expression in normal samples than in tumor cites. The endogenous expression of miR-106b was inversely correlated with MMP2 expression in MCF-7 and SUM1315-bo cells. (b) Expression of MMP2 decreased in SUM1315-bo cells after transfection of miR-106b mimic (miR-106b) compared with negative control (miR-NC) but increased in MCF-7 cells after transfection with miR-106b specific inhibitor (miR-106bI) compared with negative control (miR-NCI). (c, and d) The miR-106b binding site of MMP2 3′UTR was confirmed by luciferase assay on SUM1315-bo (c) and MCF7 (d) cells by cotransfection with the indicated reporters and miR-106b mimic or with the indicated reporters and miR-106b inhibitor. Data represent the mean ± standard deviation (SD) of at least three independent experiments. *P < 0.05, **P < 0.01.
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fig02: Matrix metalloproteinase 2 (MMP2) is the direct target of miR-106b. (a) Real-time polymerase chain reaction (PCR) analysis demonstrated higher levels of miR-106b expression in normal samples than in tumor cites. The endogenous expression of miR-106b was inversely correlated with MMP2 expression in MCF-7 and SUM1315-bo cells. (b) Expression of MMP2 decreased in SUM1315-bo cells after transfection of miR-106b mimic (miR-106b) compared with negative control (miR-NC) but increased in MCF-7 cells after transfection with miR-106b specific inhibitor (miR-106bI) compared with negative control (miR-NCI). (c, and d) The miR-106b binding site of MMP2 3′UTR was confirmed by luciferase assay on SUM1315-bo (c) and MCF7 (d) cells by cotransfection with the indicated reporters and miR-106b mimic or with the indicated reporters and miR-106b inhibitor. Data represent the mean ± standard deviation (SD) of at least three independent experiments. *P < 0.05, **P < 0.01.

Mentions: Bioinformatic analysis indicated the presence of a conserved binding site for miR-106b in MMP2 3′UTR (Suppl. Fig. S3). We found the expression level of miR-106b at the cancer site of BC patients to be lower than at the normal site (Fig.2a). The expression level of miR-106b was found to be inversely correlated with MMP2 in MCF-7 and SUM1315-bo cells. MCF-7 cells expressed more miR-106b but less MMP2 than SUM1315-bo cells (Fig.2a, Suppl. Fig. S2c,d). These results suggest a possible role for miR-106b in the regulation of MMP2. We therefore evaluated MMP2 as a target of miR-106b. To validate this hypothesis, we first investigated the effects of miR-106b overexpression on MMP2 mRNA and protein levels in SUM1315-bo cells using real-time PCR and Western blot analysis. As expected, transfection of SUM1315-bo cells with miR-106b significantly decreased MMP2 mRNA and protein levels (Fig.2b). We next investigated the effects of miR-106b-specific inhibitor (miR-106bI) on MMP2 mRNA and protein levels in MCF-7 cells. Again, miR-106bI significantly increased MMP2 mRNA and protein levels in MCF-7 cells (Fig.2b). We then focused on the potential of MMP2 as a target of the events through which miR-106b mediates the invasive and migratory activity of BC cells.


Downregulation of miR-106b induced breast cancer cell invasion and motility in association with overexpression of matrix metalloproteinase 2.

Ni X, Xia T, Zhao Y, Zhou W, Wu N, Liu X, Ding Q, Zha X, Sha J, Wang S - Cancer Sci. (2013)

Matrix metalloproteinase 2 (MMP2) is the direct target of miR-106b. (a) Real-time polymerase chain reaction (PCR) analysis demonstrated higher levels of miR-106b expression in normal samples than in tumor cites. The endogenous expression of miR-106b was inversely correlated with MMP2 expression in MCF-7 and SUM1315-bo cells. (b) Expression of MMP2 decreased in SUM1315-bo cells after transfection of miR-106b mimic (miR-106b) compared with negative control (miR-NC) but increased in MCF-7 cells after transfection with miR-106b specific inhibitor (miR-106bI) compared with negative control (miR-NCI). (c, and d) The miR-106b binding site of MMP2 3′UTR was confirmed by luciferase assay on SUM1315-bo (c) and MCF7 (d) cells by cotransfection with the indicated reporters and miR-106b mimic or with the indicated reporters and miR-106b inhibitor. Data represent the mean ± standard deviation (SD) of at least three independent experiments. *P < 0.05, **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4317878&req=5

fig02: Matrix metalloproteinase 2 (MMP2) is the direct target of miR-106b. (a) Real-time polymerase chain reaction (PCR) analysis demonstrated higher levels of miR-106b expression in normal samples than in tumor cites. The endogenous expression of miR-106b was inversely correlated with MMP2 expression in MCF-7 and SUM1315-bo cells. (b) Expression of MMP2 decreased in SUM1315-bo cells after transfection of miR-106b mimic (miR-106b) compared with negative control (miR-NC) but increased in MCF-7 cells after transfection with miR-106b specific inhibitor (miR-106bI) compared with negative control (miR-NCI). (c, and d) The miR-106b binding site of MMP2 3′UTR was confirmed by luciferase assay on SUM1315-bo (c) and MCF7 (d) cells by cotransfection with the indicated reporters and miR-106b mimic or with the indicated reporters and miR-106b inhibitor. Data represent the mean ± standard deviation (SD) of at least three independent experiments. *P < 0.05, **P < 0.01.
Mentions: Bioinformatic analysis indicated the presence of a conserved binding site for miR-106b in MMP2 3′UTR (Suppl. Fig. S3). We found the expression level of miR-106b at the cancer site of BC patients to be lower than at the normal site (Fig.2a). The expression level of miR-106b was found to be inversely correlated with MMP2 in MCF-7 and SUM1315-bo cells. MCF-7 cells expressed more miR-106b but less MMP2 than SUM1315-bo cells (Fig.2a, Suppl. Fig. S2c,d). These results suggest a possible role for miR-106b in the regulation of MMP2. We therefore evaluated MMP2 as a target of miR-106b. To validate this hypothesis, we first investigated the effects of miR-106b overexpression on MMP2 mRNA and protein levels in SUM1315-bo cells using real-time PCR and Western blot analysis. As expected, transfection of SUM1315-bo cells with miR-106b significantly decreased MMP2 mRNA and protein levels (Fig.2b). We next investigated the effects of miR-106b-specific inhibitor (miR-106bI) on MMP2 mRNA and protein levels in MCF-7 cells. Again, miR-106bI significantly increased MMP2 mRNA and protein levels in MCF-7 cells (Fig.2b). We then focused on the potential of MMP2 as a target of the events through which miR-106b mediates the invasive and migratory activity of BC cells.

Bottom Line: MMP2 was shown to be a direct target of miR-106b.Both gain- and loss-of-function studies showed that MMP2 could promote the migration and invasion of BC cells and that miR-106b could suppress both.The blockage of MMP2 by RNA interference mimicked the anti-migration and anti-invasion effects of miR-106b, and introduction of MMP2 antagonized the function of miR-106b.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China; State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, China.

Show MeSH
Related in: MedlinePlus