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Cleavage of E-Cadherin by Matrix Metalloproteinase-7 Promotes Cellular Proliferation in Nontransformed Cell Lines via Activation of RhoA.

Lynch CC, Vargo-Gogola T, Matrisian LM, Fingleton B - J Oncol (2010)

Bottom Line: Previously, we demonstrated that MMP-7, a protease implicated in mammary and intestinal tumor growth, can process the adherens junction component E-cadherin.This observation leads us to test whether MMP-7 processing of E-cadherin could directly impact cell proliferation in nontransformed epithelial cell lines (MDCK and C57MG).Our goal was to investigate the possibility that MMP-7 produced by cancer cells may have effects on adjacent normal epithelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Vanderbilt University School of Medicine, 734 PRB 2220 Pierce Ave, Nashville, TN 37232, USA.

ABSTRACT
Perturbations in cell-cell contact machinery occur frequently in epithelial cancers and result in increased cancer cell migration and invasion. Previously, we demonstrated that MMP-7, a protease implicated in mammary and intestinal tumor growth, can process the adherens junction component E-cadherin. This observation leads us to test whether MMP-7 processing of E-cadherin could directly impact cell proliferation in nontransformed epithelial cell lines (MDCK and C57MG). Our goal was to investigate the possibility that MMP-7 produced by cancer cells may have effects on adjacent normal epithelium. Here, we show that MMP-7 processing of E-cadherin mediates, (1) loss of cell-cell contact, (2) increased cell migration, (3) a loss of epithelial cell polarization and (4) increased cell proliferation via RhoA activation. These data demonstrate that MMP-7 promotes epithelial cell proliferation via the processing of E-cadherin and provide insights into the molecular mechanisms that govern epithelial cell growth.

No MeSH data available.


Related in: MedlinePlus

Epithelial proliferation is mediated by MMP-7 processing of E-cadherin. (a) Growth of the C57MG cell lines in the presence or absence of the broad spectrum metalloproteinase inhibitor (BB-94) over a 24-hour period was determined by MTT assay. Asterisk denotes that the proliferation of the MMP-7 expressing cell lines (M14/M37) is significantly higher than the empty vector controls (L2/L4), P < .05 while double asterisks denote that proliferation is significantly inhibited in the presence of BB-94 compared to the control conditions, P < .05. Significance was determined using ANOVA followed by Tukey's post hoc test. (b) The effect of E-cadherin on cell growth in the absence (IgG) or presence of an E-cadherin blocking antibody (α-E-cad) was measured using an MTT assay. Asterisk denotes significantly higher growth in the presence of the E-cadherin blocking antibody in comparison to the IgG treated controls, P < .05.  Significance was determined using ANOVA followed by Tukey's post hoc test.
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fig5: Epithelial proliferation is mediated by MMP-7 processing of E-cadherin. (a) Growth of the C57MG cell lines in the presence or absence of the broad spectrum metalloproteinase inhibitor (BB-94) over a 24-hour period was determined by MTT assay. Asterisk denotes that the proliferation of the MMP-7 expressing cell lines (M14/M37) is significantly higher than the empty vector controls (L2/L4), P < .05 while double asterisks denote that proliferation is significantly inhibited in the presence of BB-94 compared to the control conditions, P < .05. Significance was determined using ANOVA followed by Tukey's post hoc test. (b) The effect of E-cadherin on cell growth in the absence (IgG) or presence of an E-cadherin blocking antibody (α-E-cad) was measured using an MTT assay. Asterisk denotes significantly higher growth in the presence of the E-cadherin blocking antibody in comparison to the IgG treated controls, P < .05. Significance was determined using ANOVA followed by Tukey's post hoc test.

Mentions: Collectively, our data demonstrate that MMP-7 promotes disruption of the adherens junction that in turn results in a loss of epithelial polarization and increased proliferation. Given that MMP-7 can mediate E-cadherin processing, we next examined whether the effects on epithelial cell proliferation were related to the E-cadherin cleavage. We first confirmed that the proliferation phenotype directly resulted from metalloproteinase activity. To this end, the empty vector and MMP-7 expressing C57MG clones were incubated in the presence or absence of the metalloproteinase inhibitor, BB-94. Over 24 hours, we identified that treatment with BB-94 (5 μM) reduced the proliferation of the MMP-7 expressing cell lines to the level of the control empty vector cell lines suggesting that metalloproteinases could promote the proliferation of nontransformed mammary epithelial cells directly as assessed by MTT assay (L2 Control: 1.74 ± 0.25 versus L2 BB-94 5 μM: 1.8 ± 0.15, L4 Control: 2.04 ± 0.13 versus L4 BB-94 5 μM: 2.01 ± 0.15, M14 Control: 2.51 ± 0.27 versus M14 BB-94 5 μM: 1.993 ± 0.06, M37 Control 2.75 ± 0.21 versus M37 BB-94 5 μM: 2.237 ± 0.10 ABS at 490 nm, Figure 5(a)). Of note, BB-94 did not impact the proliferative ability of the empty vector control cell lines (Figure 5(a)), thus suggesting that MMP-7 is the metalloproteinase promoting the proliferative phenotype in these cells.


Cleavage of E-Cadherin by Matrix Metalloproteinase-7 Promotes Cellular Proliferation in Nontransformed Cell Lines via Activation of RhoA.

Lynch CC, Vargo-Gogola T, Matrisian LM, Fingleton B - J Oncol (2010)

Epithelial proliferation is mediated by MMP-7 processing of E-cadherin. (a) Growth of the C57MG cell lines in the presence or absence of the broad spectrum metalloproteinase inhibitor (BB-94) over a 24-hour period was determined by MTT assay. Asterisk denotes that the proliferation of the MMP-7 expressing cell lines (M14/M37) is significantly higher than the empty vector controls (L2/L4), P < .05 while double asterisks denote that proliferation is significantly inhibited in the presence of BB-94 compared to the control conditions, P < .05. Significance was determined using ANOVA followed by Tukey's post hoc test. (b) The effect of E-cadherin on cell growth in the absence (IgG) or presence of an E-cadherin blocking antibody (α-E-cad) was measured using an MTT assay. Asterisk denotes significantly higher growth in the presence of the E-cadherin blocking antibody in comparison to the IgG treated controls, P < .05.  Significance was determined using ANOVA followed by Tukey's post hoc test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Epithelial proliferation is mediated by MMP-7 processing of E-cadherin. (a) Growth of the C57MG cell lines in the presence or absence of the broad spectrum metalloproteinase inhibitor (BB-94) over a 24-hour period was determined by MTT assay. Asterisk denotes that the proliferation of the MMP-7 expressing cell lines (M14/M37) is significantly higher than the empty vector controls (L2/L4), P < .05 while double asterisks denote that proliferation is significantly inhibited in the presence of BB-94 compared to the control conditions, P < .05. Significance was determined using ANOVA followed by Tukey's post hoc test. (b) The effect of E-cadherin on cell growth in the absence (IgG) or presence of an E-cadherin blocking antibody (α-E-cad) was measured using an MTT assay. Asterisk denotes significantly higher growth in the presence of the E-cadherin blocking antibody in comparison to the IgG treated controls, P < .05. Significance was determined using ANOVA followed by Tukey's post hoc test.
Mentions: Collectively, our data demonstrate that MMP-7 promotes disruption of the adherens junction that in turn results in a loss of epithelial polarization and increased proliferation. Given that MMP-7 can mediate E-cadherin processing, we next examined whether the effects on epithelial cell proliferation were related to the E-cadherin cleavage. We first confirmed that the proliferation phenotype directly resulted from metalloproteinase activity. To this end, the empty vector and MMP-7 expressing C57MG clones were incubated in the presence or absence of the metalloproteinase inhibitor, BB-94. Over 24 hours, we identified that treatment with BB-94 (5 μM) reduced the proliferation of the MMP-7 expressing cell lines to the level of the control empty vector cell lines suggesting that metalloproteinases could promote the proliferation of nontransformed mammary epithelial cells directly as assessed by MTT assay (L2 Control: 1.74 ± 0.25 versus L2 BB-94 5 μM: 1.8 ± 0.15, L4 Control: 2.04 ± 0.13 versus L4 BB-94 5 μM: 2.01 ± 0.15, M14 Control: 2.51 ± 0.27 versus M14 BB-94 5 μM: 1.993 ± 0.06, M37 Control 2.75 ± 0.21 versus M37 BB-94 5 μM: 2.237 ± 0.10 ABS at 490 nm, Figure 5(a)). Of note, BB-94 did not impact the proliferative ability of the empty vector control cell lines (Figure 5(a)), thus suggesting that MMP-7 is the metalloproteinase promoting the proliferative phenotype in these cells.

Bottom Line: Previously, we demonstrated that MMP-7, a protease implicated in mammary and intestinal tumor growth, can process the adherens junction component E-cadherin.This observation leads us to test whether MMP-7 processing of E-cadherin could directly impact cell proliferation in nontransformed epithelial cell lines (MDCK and C57MG).Our goal was to investigate the possibility that MMP-7 produced by cancer cells may have effects on adjacent normal epithelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Vanderbilt University School of Medicine, 734 PRB 2220 Pierce Ave, Nashville, TN 37232, USA.

ABSTRACT
Perturbations in cell-cell contact machinery occur frequently in epithelial cancers and result in increased cancer cell migration and invasion. Previously, we demonstrated that MMP-7, a protease implicated in mammary and intestinal tumor growth, can process the adherens junction component E-cadherin. This observation leads us to test whether MMP-7 processing of E-cadherin could directly impact cell proliferation in nontransformed epithelial cell lines (MDCK and C57MG). Our goal was to investigate the possibility that MMP-7 produced by cancer cells may have effects on adjacent normal epithelium. Here, we show that MMP-7 processing of E-cadherin mediates, (1) loss of cell-cell contact, (2) increased cell migration, (3) a loss of epithelial cell polarization and (4) increased cell proliferation via RhoA activation. These data demonstrate that MMP-7 promotes epithelial cell proliferation via the processing of E-cadherin and provide insights into the molecular mechanisms that govern epithelial cell growth.

No MeSH data available.


Related in: MedlinePlus