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Immunolocalization of MMP9 and MMP2 in osteolytic metastasis originating from MDA-MB-231 human breast cancer cells.

Liu B, Cui J, Sun J, Li J, Han X, Guo J, Yi M, Amizuka N, Xu X, Li M - Mol Med Rep (2016)

Bottom Line: Numerous tartrate-resistant acid phosphatase-positive osteoclasts were found in the metastasized lesions.The invaded tumor cells positive for mammaglobin 1 exhibited different proliferation activities and apoptosis between the metaphysis and diaphysis.Of note, MMP9 was expressed predominantly in the proliferating cell nuclear antigen‑positive area, whereas the expression of MMP2 was observed predominantly in the diaphysis, which had more TUNEL‑positive cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Bone Metabolism, School of Stomatology, Shandong University, Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan, Shandong 250012, P.R. China.

ABSTRACT
The aim of the present study was to investigate the expression of matrix metalloproteinase (MMP)9 and MMP2, and their potential roles in bone metastasis nests using a well-standardized model of breast cancer bone metastasis in nude mice. BALB/c nu/nu mice (5-week-old; n=10) were subjected to intracardiac injection of MDA-MB-231 human breast cancer cells. After 4 weeks, the mice exhibiting radiolucent lesions in tibiae were sacrificed, and the tibiae were removed for histochemical analysis. The gene expression of MMP2 and MMP9 in the tumor cells, metaphysis and diaphysis of normal BALB/c nu/nu mice were determined using reverse transcription-polymerase chain reaction analysis. The metastatic tumor tissue occupied almost the entire bone marrow cavity. Numerous tartrate-resistant acid phosphatase-positive osteoclasts were found in the metastasized lesions. The invaded tumor cells positive for mammaglobin 1 exhibited different proliferation activities and apoptosis between the metaphysis and diaphysis. Proliferating cell nuclear antigen was expressed at high levels in the metaphyseal area, whereas TdT-mediated dUTP nick-end labeling (TUNEL)-positive cells were more evident in the diaphysis area. Of note, MMP9 was expressed predominantly in the proliferating cell nuclear antigen‑positive area, whereas the expression of MMP2 was observed predominantly in the diaphysis, which had more TUNEL‑positive cells. Taken together, the results suggested that MMP9 and MMP2 may have their own importance in extracellular matrix degradation and trabecular bone damage in different zones of bone metastasis, including the metaphysis and diaphysis.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemical and statistical analyses of MMP9 and MMP2. (A) Immunohistochemical analysis of MMP9 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP9 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP9 was significantly higher in the metaphysis, compared with the diaphysis. (e and f) High magnification of c and d, respectively. (B) Immunohistochemical analysis of MMP2 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP2 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP2 was significantly higher in the diaphysis, compared with the metaphysis. (e and f) High magnification of c and d, respectively. (C) Statistical analyses of the immunostaining intensity of MMP9 and MMP2 in the metaphysis and diaphysis. *P<0.01. Scale bars=50 µm in Aa-d and Ba-d; 25 µm in Ae and f, and Be and B. MMP, matrix metalloproteinase; Meta, metaphysis; Dia, diaphysis; T, tumor cells; CB, cortical bone; *, normal bone tissue.
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f3-mmr-14-02-1099: Immunohistochemical and statistical analyses of MMP9 and MMP2. (A) Immunohistochemical analysis of MMP9 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP9 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP9 was significantly higher in the metaphysis, compared with the diaphysis. (e and f) High magnification of c and d, respectively. (B) Immunohistochemical analysis of MMP2 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP2 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP2 was significantly higher in the diaphysis, compared with the metaphysis. (e and f) High magnification of c and d, respectively. (C) Statistical analyses of the immunostaining intensity of MMP9 and MMP2 in the metaphysis and diaphysis. *P<0.01. Scale bars=50 µm in Aa-d and Ba-d; 25 µm in Ae and f, and Be and B. MMP, matrix metalloproteinase; Meta, metaphysis; Dia, diaphysis; T, tumor cells; CB, cortical bone; *, normal bone tissue.

Mentions: In the metaphyseal area containing numerous PCNA-positive tumor cells, MMP9-immunopositivity was significantly more marked (Fig. 3Ab and c), whereas staining for MMP2 was faint (Fig. 3Bb and c). In contrast, the diaphyseal metastasis, containing more TUNEL-positive cells, showed weak expression of MMP9 (Fig. 3Ae and f), compared with MMP2 (Fig. 3Be and f). No significant differences were found in the immunolocalization and immunoreactivity of MMP13 between PCNA-positive and negative areas (data not shown). ANOVA revealed that, in the PCNA-positive metastatic area (metaphysis), the staining intensity of MMP2 was significantly weaker, compared with that of MMP9 (0.126±0.007, vs. 0.300±0.036, respectively; P<0.01; n=7; Fig. 3C). In the TUNEL-positive metastatic area (diaphysis), the staining intensity of MMP2 was significantly more marked, compared with that of MMP9 (0.205±0.020, vs. 0.103±0.009; P<0.01; Fig. 3C).


Immunolocalization of MMP9 and MMP2 in osteolytic metastasis originating from MDA-MB-231 human breast cancer cells.

Liu B, Cui J, Sun J, Li J, Han X, Guo J, Yi M, Amizuka N, Xu X, Li M - Mol Med Rep (2016)

Immunohistochemical and statistical analyses of MMP9 and MMP2. (A) Immunohistochemical analysis of MMP9 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP9 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP9 was significantly higher in the metaphysis, compared with the diaphysis. (e and f) High magnification of c and d, respectively. (B) Immunohistochemical analysis of MMP2 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP2 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP2 was significantly higher in the diaphysis, compared with the metaphysis. (e and f) High magnification of c and d, respectively. (C) Statistical analyses of the immunostaining intensity of MMP9 and MMP2 in the metaphysis and diaphysis. *P<0.01. Scale bars=50 µm in Aa-d and Ba-d; 25 µm in Ae and f, and Be and B. MMP, matrix metalloproteinase; Meta, metaphysis; Dia, diaphysis; T, tumor cells; CB, cortical bone; *, normal bone tissue.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4940081&req=5

f3-mmr-14-02-1099: Immunohistochemical and statistical analyses of MMP9 and MMP2. (A) Immunohistochemical analysis of MMP9 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP9 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP9 was significantly higher in the metaphysis, compared with the diaphysis. (e and f) High magnification of c and d, respectively. (B) Immunohistochemical analysis of MMP2 in the (a) metaphysis and (b) diaphysis of normal bone marrow in the control group. Immunohistochemical analysis of MMP2 in tumor tissue of the (c) metaphysis and (d) diaphysis (brown color). The expression of MMP2 was significantly higher in the diaphysis, compared with the metaphysis. (e and f) High magnification of c and d, respectively. (C) Statistical analyses of the immunostaining intensity of MMP9 and MMP2 in the metaphysis and diaphysis. *P<0.01. Scale bars=50 µm in Aa-d and Ba-d; 25 µm in Ae and f, and Be and B. MMP, matrix metalloproteinase; Meta, metaphysis; Dia, diaphysis; T, tumor cells; CB, cortical bone; *, normal bone tissue.
Mentions: In the metaphyseal area containing numerous PCNA-positive tumor cells, MMP9-immunopositivity was significantly more marked (Fig. 3Ab and c), whereas staining for MMP2 was faint (Fig. 3Bb and c). In contrast, the diaphyseal metastasis, containing more TUNEL-positive cells, showed weak expression of MMP9 (Fig. 3Ae and f), compared with MMP2 (Fig. 3Be and f). No significant differences were found in the immunolocalization and immunoreactivity of MMP13 between PCNA-positive and negative areas (data not shown). ANOVA revealed that, in the PCNA-positive metastatic area (metaphysis), the staining intensity of MMP2 was significantly weaker, compared with that of MMP9 (0.126±0.007, vs. 0.300±0.036, respectively; P<0.01; n=7; Fig. 3C). In the TUNEL-positive metastatic area (diaphysis), the staining intensity of MMP2 was significantly more marked, compared with that of MMP9 (0.205±0.020, vs. 0.103±0.009; P<0.01; Fig. 3C).

Bottom Line: Numerous tartrate-resistant acid phosphatase-positive osteoclasts were found in the metastasized lesions.The invaded tumor cells positive for mammaglobin 1 exhibited different proliferation activities and apoptosis between the metaphysis and diaphysis.Of note, MMP9 was expressed predominantly in the proliferating cell nuclear antigen‑positive area, whereas the expression of MMP2 was observed predominantly in the diaphysis, which had more TUNEL‑positive cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Bone Metabolism, School of Stomatology, Shandong University, Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan, Shandong 250012, P.R. China.

ABSTRACT
The aim of the present study was to investigate the expression of matrix metalloproteinase (MMP)9 and MMP2, and their potential roles in bone metastasis nests using a well-standardized model of breast cancer bone metastasis in nude mice. BALB/c nu/nu mice (5-week-old; n=10) were subjected to intracardiac injection of MDA-MB-231 human breast cancer cells. After 4 weeks, the mice exhibiting radiolucent lesions in tibiae were sacrificed, and the tibiae were removed for histochemical analysis. The gene expression of MMP2 and MMP9 in the tumor cells, metaphysis and diaphysis of normal BALB/c nu/nu mice were determined using reverse transcription-polymerase chain reaction analysis. The metastatic tumor tissue occupied almost the entire bone marrow cavity. Numerous tartrate-resistant acid phosphatase-positive osteoclasts were found in the metastasized lesions. The invaded tumor cells positive for mammaglobin 1 exhibited different proliferation activities and apoptosis between the metaphysis and diaphysis. Proliferating cell nuclear antigen was expressed at high levels in the metaphyseal area, whereas TdT-mediated dUTP nick-end labeling (TUNEL)-positive cells were more evident in the diaphysis area. Of note, MMP9 was expressed predominantly in the proliferating cell nuclear antigen‑positive area, whereas the expression of MMP2 was observed predominantly in the diaphysis, which had more TUNEL‑positive cells. Taken together, the results suggested that MMP9 and MMP2 may have their own importance in extracellular matrix degradation and trabecular bone damage in different zones of bone metastasis, including the metaphysis and diaphysis.

No MeSH data available.


Related in: MedlinePlus