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Pleiotropic functions of the tumor- and metastasis-suppressing matrix metalloproteinase-8 in mammary cancer in MMTV-PyMT transgenic mice.

Decock J, Hendrickx W, Thirkettle S, Gutiérrez-Fernández A, Robinson SD, Edwards DR - Breast Cancer Res. (2015)

Bottom Line: No differences in macrophage infiltration were apparent throughout primary tumor development, though at 10 weeks a drop in neutrophil infiltrates was observed in Mmp8-wild-type tumors.Using quantitative real-time RT-PCR, we tracked the expression of the entire Mmp and Timp gene families, observing a significant decrease in Mmp3 expression in Mmp8- tumors compared to wild-type and heterozygotes throughout the time course of the model, which was confirmed at the protein level.These findings provide novel insight into the suppressive action of MMP-8 on mammary tumorigenesis and metastasis, and indicate that the loss of MMP-8 likely has pleiotropic effects on innate immunity and angiogenesis that are reflected in changes in the protease web.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Matrix metalloproteinase-8 (MMP-8; neutrophil collagenase) is an important regulator of innate immunity that has oncosuppressive actions in numerous tumor types.

Methods: We have intercrossed Mmp8- mice with the Polyoma virus middle T oncogene-driven (MMTV-PyMT) mouse model of mammary cancer to explore the effects of loss of MMP-8 on the incidence and progression of mammary carcinomas.

Results: In this aggressive mouse model of breast cancer, loss of MMP-8 accelerated tumor onset even further, such that 90% of MMTV-PyMT; Mmp8- female mice were tumor-bearing at the time of weaning. Throughout the 14 weeks of the model, tumor burden increased in homozygous Mmp8- mice compared to Mmp8-wild-type and -heterozygote animals. Likewise, lung metastasis dramatically increased in the MMTV-PyMT; Mmp8- mice. Immunohistochemistry revealed that tumors in wild-type, Mmp8-heterozygotes and - animals had similar vascular density at 8 weeks, but at 10 weeks Mmp8-wild-type tumors had a lower vascularity than their heterozygote and counterparts. No differences in macrophage infiltration were apparent throughout primary tumor development, though at 10 weeks a drop in neutrophil infiltrates was observed in Mmp8-wild-type tumors. Using quantitative real-time RT-PCR, we tracked the expression of the entire Mmp and Timp gene families, observing a significant decrease in Mmp3 expression in Mmp8- tumors compared to wild-type and heterozygotes throughout the time course of the model, which was confirmed at the protein level.

Conclusions: These findings provide novel insight into the suppressive action of MMP-8 on mammary tumorigenesis and metastasis, and indicate that the loss of MMP-8 likely has pleiotropic effects on innate immunity and angiogenesis that are reflected in changes in the protease web.

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Loss of MMP-8 accelerates tumor onset; promotes progression, tumor size and lung macrometastases in the MMTV-PyMT model. (A) Detection of first palpable tumors of MMTV-PyMT; Mmp8-wild-type (WT, n = 17), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 10) mice. Mean ± SEM, two-tailed unpaired t test; *P <0.05; **P <0.01. (B) Tumor progression determined by palpation of all 10 mammary glands from day 21 to day 98 for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), −heterozygote (HET, n = 34) and - (KO, n = 21) mice. (C) Tumor size distribution (0, <0.5, 0.5 to 1, >1 cm) at 10 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), MMTV-PyMT; Mmp8-heterozygote (HET, n = 34) and MMTV-PyMT; Mmp8- (KO, n = 21) mice; and at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 22), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 16) mice. chi-square test, *P <0.01. (D) Number of lung macrometastases assessed at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 4), MMTV-PyMT; Mmp8-heterozygote (HET, n = 4) and MMTV-PyMT; Mmp8- (KO, n = 2) mice. Left panel of representative pictures depicts the top and bottom of all lung lobes (one lobe of the left lung, four lobes of the right lung). Black solid arrows = smaller macrometastases, white solid arrows = larger macrometastases. Mean ± SEM, two-tailed unpaired t test; **P <0.01. MMP, matrix metalloproteinase; MMTV, mouse mammary tumor virus; PyMT, Polyoma virus middle T-antigen; SEM, standard error of the mean.
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Fig1: Loss of MMP-8 accelerates tumor onset; promotes progression, tumor size and lung macrometastases in the MMTV-PyMT model. (A) Detection of first palpable tumors of MMTV-PyMT; Mmp8-wild-type (WT, n = 17), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 10) mice. Mean ± SEM, two-tailed unpaired t test; *P <0.05; **P <0.01. (B) Tumor progression determined by palpation of all 10 mammary glands from day 21 to day 98 for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), −heterozygote (HET, n = 34) and - (KO, n = 21) mice. (C) Tumor size distribution (0, <0.5, 0.5 to 1, >1 cm) at 10 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), MMTV-PyMT; Mmp8-heterozygote (HET, n = 34) and MMTV-PyMT; Mmp8- (KO, n = 21) mice; and at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 22), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 16) mice. chi-square test, *P <0.01. (D) Number of lung macrometastases assessed at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 4), MMTV-PyMT; Mmp8-heterozygote (HET, n = 4) and MMTV-PyMT; Mmp8- (KO, n = 2) mice. Left panel of representative pictures depicts the top and bottom of all lung lobes (one lobe of the left lung, four lobes of the right lung). Black solid arrows = smaller macrometastases, white solid arrows = larger macrometastases. Mean ± SEM, two-tailed unpaired t test; **P <0.01. MMP, matrix metalloproteinase; MMTV, mouse mammary tumor virus; PyMT, Polyoma virus middle T-antigen; SEM, standard error of the mean.

Mentions: The effect of MMP-8 ablation on tumor onset and growth was investigated in cohorts of Mmp8-wild-type, heterozygote and female PyMT littermates [27]. All 10 mammary glands were palpated biweekly for the presence of tumors. In MMTV-PyMT; Mmp8- mice (KO; n = 10), tumors were first detected at the age of 25 days compared to 32 days in wild-type (WT; n = 17, P <0.01) and heterozygote mice (HET; n = 30, P <0.05) (Figure 1A). All MMTV-PyMT; Mmp8- mice (n = 21) were tumor-bearing at the age of 40 days, whereas full penetrance in heterozygote (n = 34) and wild-type (n = 26) mice was not observed until 50 and 55 days respectively (Figure 1B). No difference in tumor multiplicity was found (data not shown).Figure 1


Pleiotropic functions of the tumor- and metastasis-suppressing matrix metalloproteinase-8 in mammary cancer in MMTV-PyMT transgenic mice.

Decock J, Hendrickx W, Thirkettle S, Gutiérrez-Fernández A, Robinson SD, Edwards DR - Breast Cancer Res. (2015)

Loss of MMP-8 accelerates tumor onset; promotes progression, tumor size and lung macrometastases in the MMTV-PyMT model. (A) Detection of first palpable tumors of MMTV-PyMT; Mmp8-wild-type (WT, n = 17), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 10) mice. Mean ± SEM, two-tailed unpaired t test; *P <0.05; **P <0.01. (B) Tumor progression determined by palpation of all 10 mammary glands from day 21 to day 98 for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), −heterozygote (HET, n = 34) and - (KO, n = 21) mice. (C) Tumor size distribution (0, <0.5, 0.5 to 1, >1 cm) at 10 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), MMTV-PyMT; Mmp8-heterozygote (HET, n = 34) and MMTV-PyMT; Mmp8- (KO, n = 21) mice; and at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 22), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 16) mice. chi-square test, *P <0.01. (D) Number of lung macrometastases assessed at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 4), MMTV-PyMT; Mmp8-heterozygote (HET, n = 4) and MMTV-PyMT; Mmp8- (KO, n = 2) mice. Left panel of representative pictures depicts the top and bottom of all lung lobes (one lobe of the left lung, four lobes of the right lung). Black solid arrows = smaller macrometastases, white solid arrows = larger macrometastases. Mean ± SEM, two-tailed unpaired t test; **P <0.01. MMP, matrix metalloproteinase; MMTV, mouse mammary tumor virus; PyMT, Polyoma virus middle T-antigen; SEM, standard error of the mean.
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Fig1: Loss of MMP-8 accelerates tumor onset; promotes progression, tumor size and lung macrometastases in the MMTV-PyMT model. (A) Detection of first palpable tumors of MMTV-PyMT; Mmp8-wild-type (WT, n = 17), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 10) mice. Mean ± SEM, two-tailed unpaired t test; *P <0.05; **P <0.01. (B) Tumor progression determined by palpation of all 10 mammary glands from day 21 to day 98 for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), −heterozygote (HET, n = 34) and - (KO, n = 21) mice. (C) Tumor size distribution (0, <0.5, 0.5 to 1, >1 cm) at 10 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 26), MMTV-PyMT; Mmp8-heterozygote (HET, n = 34) and MMTV-PyMT; Mmp8- (KO, n = 21) mice; and at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 22), MMTV-PyMT; Mmp8-heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8- (KO, n = 16) mice. chi-square test, *P <0.01. (D) Number of lung macrometastases assessed at 14 weeks of age for MMTV-PyMT; Mmp8-wild-type (WT, n = 4), MMTV-PyMT; Mmp8-heterozygote (HET, n = 4) and MMTV-PyMT; Mmp8- (KO, n = 2) mice. Left panel of representative pictures depicts the top and bottom of all lung lobes (one lobe of the left lung, four lobes of the right lung). Black solid arrows = smaller macrometastases, white solid arrows = larger macrometastases. Mean ± SEM, two-tailed unpaired t test; **P <0.01. MMP, matrix metalloproteinase; MMTV, mouse mammary tumor virus; PyMT, Polyoma virus middle T-antigen; SEM, standard error of the mean.
Mentions: The effect of MMP-8 ablation on tumor onset and growth was investigated in cohorts of Mmp8-wild-type, heterozygote and female PyMT littermates [27]. All 10 mammary glands were palpated biweekly for the presence of tumors. In MMTV-PyMT; Mmp8- mice (KO; n = 10), tumors were first detected at the age of 25 days compared to 32 days in wild-type (WT; n = 17, P <0.01) and heterozygote mice (HET; n = 30, P <0.05) (Figure 1A). All MMTV-PyMT; Mmp8- mice (n = 21) were tumor-bearing at the age of 40 days, whereas full penetrance in heterozygote (n = 34) and wild-type (n = 26) mice was not observed until 50 and 55 days respectively (Figure 1B). No difference in tumor multiplicity was found (data not shown).Figure 1

Bottom Line: No differences in macrophage infiltration were apparent throughout primary tumor development, though at 10 weeks a drop in neutrophil infiltrates was observed in Mmp8-wild-type tumors.Using quantitative real-time RT-PCR, we tracked the expression of the entire Mmp and Timp gene families, observing a significant decrease in Mmp3 expression in Mmp8- tumors compared to wild-type and heterozygotes throughout the time course of the model, which was confirmed at the protein level.These findings provide novel insight into the suppressive action of MMP-8 on mammary tumorigenesis and metastasis, and indicate that the loss of MMP-8 likely has pleiotropic effects on innate immunity and angiogenesis that are reflected in changes in the protease web.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Matrix metalloproteinase-8 (MMP-8; neutrophil collagenase) is an important regulator of innate immunity that has oncosuppressive actions in numerous tumor types.

Methods: We have intercrossed Mmp8- mice with the Polyoma virus middle T oncogene-driven (MMTV-PyMT) mouse model of mammary cancer to explore the effects of loss of MMP-8 on the incidence and progression of mammary carcinomas.

Results: In this aggressive mouse model of breast cancer, loss of MMP-8 accelerated tumor onset even further, such that 90% of MMTV-PyMT; Mmp8- female mice were tumor-bearing at the time of weaning. Throughout the 14 weeks of the model, tumor burden increased in homozygous Mmp8- mice compared to Mmp8-wild-type and -heterozygote animals. Likewise, lung metastasis dramatically increased in the MMTV-PyMT; Mmp8- mice. Immunohistochemistry revealed that tumors in wild-type, Mmp8-heterozygotes and - animals had similar vascular density at 8 weeks, but at 10 weeks Mmp8-wild-type tumors had a lower vascularity than their heterozygote and counterparts. No differences in macrophage infiltration were apparent throughout primary tumor development, though at 10 weeks a drop in neutrophil infiltrates was observed in Mmp8-wild-type tumors. Using quantitative real-time RT-PCR, we tracked the expression of the entire Mmp and Timp gene families, observing a significant decrease in Mmp3 expression in Mmp8- tumors compared to wild-type and heterozygotes throughout the time course of the model, which was confirmed at the protein level.

Conclusions: These findings provide novel insight into the suppressive action of MMP-8 on mammary tumorigenesis and metastasis, and indicate that the loss of MMP-8 likely has pleiotropic effects on innate immunity and angiogenesis that are reflected in changes in the protease web.

Show MeSH
Related in: MedlinePlus