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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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Effects of matrix metalloproteinase 2 (MMP2) on migration and invasion of breast cancer (BC) cells. (a) The siRNA of MMP2 was transfected into SUM1315-bo. This was confirmed at both the gene and protein levels. Si-M1, si-M2, and si-M3 represent the three different siRNA pairs of MMP2. Si-M1 and si-M2 were used in all the subsequent experiments. (b) The invasiveness of MMP2 knockdown SUM1315-bo and the negative control (si-NC) cells were assessed using transwell assays. The invasiveness through 8 μm pore transwells was significantly lower in MMP2 knockdown SUM1315-bo than in the negative control (P < 0.05). Original magnification ×100. (c) The constructed expression vector pEGFP-C2-MMP2 was stably transfected into MCF-7, which was MMP2-negative. Transfection was confirmed at the protein level. (d) The invasiveness of MMP2 overexpressing MCF-7 (MCF7-MO) and the control cells were assessed by transwell assays. The invasiveness through 8 μm pore transwells was found to be significantly higher in MMP2 over-expressing MCF-7 than in controls (P < 0.05). Original magnification ×100.
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fig01: Effects of matrix metalloproteinase 2 (MMP2) on migration and invasion of breast cancer (BC) cells. (a) The siRNA of MMP2 was transfected into SUM1315-bo. This was confirmed at both the gene and protein levels. Si-M1, si-M2, and si-M3 represent the three different siRNA pairs of MMP2. Si-M1 and si-M2 were used in all the subsequent experiments. (b) The invasiveness of MMP2 knockdown SUM1315-bo and the negative control (si-NC) cells were assessed using transwell assays. The invasiveness through 8 μm pore transwells was significantly lower in MMP2 knockdown SUM1315-bo than in the negative control (P < 0.05). Original magnification ×100. (c) The constructed expression vector pEGFP-C2-MMP2 was stably transfected into MCF-7, which was MMP2-negative. Transfection was confirmed at the protein level. (d) The invasiveness of MMP2 overexpressing MCF-7 (MCF7-MO) and the control cells were assessed by transwell assays. The invasiveness through 8 μm pore transwells was found to be significantly higher in MMP2 over-expressing MCF-7 than in controls (P < 0.05). Original magnification ×100.

Mentions: The role of MMP2 on cell migration and invasion of SUM1315-bo cells was demonstrated by knocking down MMP2 using siRNA. We used SUM1315-bo cells because they have a high migratory potential and express endogenous MMP2 in large quantities. Three small interfering RNA (siRNA)-coding oligos against human MMP2 were designed and compared. The two most effective MMP2 siRNA (si-M1 and si-M2) constructs had target sequences of AGU AGA UCC AGU AUU CAU UCC CUG C (si-M1) and AGA AGU UGU AGU UGG CCA CAU CUG G (si-M2). These sequences were verified at the mRNA and protein levels (Fig.1a). Notably, silencing of MMP2 was found to be associated with significantly decreased invasive and migratory ability in SUM1315-bo cells (Fig.1b).


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)

Effects of matrix metalloproteinase 2 (MMP2) on migration and invasion of breast cancer (BC) cells. (a) The siRNA of MMP2 was transfected into SUM1315-bo. This was confirmed at both the gene and protein levels. Si-M1, si-M2, and si-M3 represent the three different siRNA pairs of MMP2. Si-M1 and si-M2 were used in all the subsequent experiments. (b) The invasiveness of MMP2 knockdown SUM1315-bo and the negative control (si-NC) cells were assessed using transwell assays. The invasiveness through 8 μm pore transwells was significantly lower in MMP2 knockdown SUM1315-bo than in the negative control (P < 0.05). Original magnification ×100. (c) The constructed expression vector pEGFP-C2-MMP2 was stably transfected into MCF-7, which was MMP2-negative. Transfection was confirmed at the protein level. (d) The invasiveness of MMP2 overexpressing MCF-7 (MCF7-MO) and the control cells were assessed by transwell assays. The invasiveness through 8 μm pore transwells was found to be significantly higher in MMP2 over-expressing MCF-7 than in controls (P < 0.05). Original magnification ×100.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317878&req=5

fig01: Effects of matrix metalloproteinase 2 (MMP2) on migration and invasion of breast cancer (BC) cells. (a) The siRNA of MMP2 was transfected into SUM1315-bo. This was confirmed at both the gene and protein levels. Si-M1, si-M2, and si-M3 represent the three different siRNA pairs of MMP2. Si-M1 and si-M2 were used in all the subsequent experiments. (b) The invasiveness of MMP2 knockdown SUM1315-bo and the negative control (si-NC) cells were assessed using transwell assays. The invasiveness through 8 μm pore transwells was significantly lower in MMP2 knockdown SUM1315-bo than in the negative control (P < 0.05). Original magnification ×100. (c) The constructed expression vector pEGFP-C2-MMP2 was stably transfected into MCF-7, which was MMP2-negative. Transfection was confirmed at the protein level. (d) The invasiveness of MMP2 overexpressing MCF-7 (MCF7-MO) and the control cells were assessed by transwell assays. The invasiveness through 8 μm pore transwells was found to be significantly higher in MMP2 over-expressing MCF-7 than in controls (P < 0.05). Original magnification ×100.
Mentions: The role of MMP2 on cell migration and invasion of SUM1315-bo cells was demonstrated by knocking down MMP2 using siRNA. We used SUM1315-bo cells because they have a high migratory potential and express endogenous MMP2 in large quantities. Three small interfering RNA (siRNA)-coding oligos against human MMP2 were designed and compared. The two most effective MMP2 siRNA (si-M1 and si-M2) constructs had target sequences of AGU AGA UCC AGU AUU CAU UCC CUG C (si-M1) and AGA AGU UGU AGU UGG CCA CAU CUG G (si-M2). These sequences were verified at the mRNA and protein levels (Fig.1a). Notably, silencing of MMP2 was found to be associated with significantly decreased invasive and migratory ability in SUM1315-bo cells (Fig.1b).

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