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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

Reverse transcription-polymerase chain reaction analysis of the mRNA expression of MMP2 and MMP9 in MDA-MB-231 cells, and the metaphysis and diaphysis of tibiae of BALB/c nu/nu mice. M, Tiangen DM:120227 DNA marker (50-500 bp) C, Human-β-actin (250 bp); C1, Mus-β-actin (154 bp); T1, target gene 1 (Human-MMP2) was positive in the MDA-MB-231 cells with a size of 112 bp). T2, target gene 2 (Human-MMP9) was positive in the MDA-MB-231 cells with a size of 139 bp). T3, target gene 3 (Mus-MMP2) was positive in the metaphysis of the tibiae of BALB/c nu/nu mice with a size of 140 bp). T4, target gene 4 (Mus-MMP9 was positive in the diaphysis of the tibiae of BALB/c nu/nu mice with a size of 229 bp. MMP, matrix metalloproteinase.
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f4-mmr-14-02-1099: Reverse transcription-polymerase chain reaction analysis of the mRNA expression of MMP2 and MMP9 in MDA-MB-231 cells, and the metaphysis and diaphysis of tibiae of BALB/c nu/nu mice. M, Tiangen DM:120227 DNA marker (50-500 bp) C, Human-β-actin (250 bp); C1, Mus-β-actin (154 bp); T1, target gene 1 (Human-MMP2) was positive in the MDA-MB-231 cells with a size of 112 bp). T2, target gene 2 (Human-MMP9) was positive in the MDA-MB-231 cells with a size of 139 bp). T3, target gene 3 (Mus-MMP2) was positive in the metaphysis of the tibiae of BALB/c nu/nu mice with a size of 140 bp). T4, target gene 4 (Mus-MMP9 was positive in the diaphysis of the tibiae of BALB/c nu/nu mice with a size of 229 bp. MMP, matrix metalloproteinase.

Mentions: The present study performed RT-PCR to investigate the source of MMP2 and MMP9. The results revealed that MMP2 and MMP9 mRNA were expressed in the MDA-MB-231 cells, metaphysis and diaphysis of the tibiae of the BALB/c nu/nu mice without tumor cell administration (Fig. 4).


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)

Reverse transcription-polymerase chain reaction analysis of the mRNA expression of MMP2 and MMP9 in MDA-MB-231 cells, and the metaphysis and diaphysis of tibiae of BALB/c nu/nu mice. M, Tiangen DM:120227 DNA marker (50-500 bp) C, Human-β-actin (250 bp); C1, Mus-β-actin (154 bp); T1, target gene 1 (Human-MMP2) was positive in the MDA-MB-231 cells with a size of 112 bp). T2, target gene 2 (Human-MMP9) was positive in the MDA-MB-231 cells with a size of 139 bp). T3, target gene 3 (Mus-MMP2) was positive in the metaphysis of the tibiae of BALB/c nu/nu mice with a size of 140 bp). T4, target gene 4 (Mus-MMP9 was positive in the diaphysis of the tibiae of BALB/c nu/nu mice with a size of 229 bp. MMP, matrix metalloproteinase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940081&req=5

f4-mmr-14-02-1099: Reverse transcription-polymerase chain reaction analysis of the mRNA expression of MMP2 and MMP9 in MDA-MB-231 cells, and the metaphysis and diaphysis of tibiae of BALB/c nu/nu mice. M, Tiangen DM:120227 DNA marker (50-500 bp) C, Human-β-actin (250 bp); C1, Mus-β-actin (154 bp); T1, target gene 1 (Human-MMP2) was positive in the MDA-MB-231 cells with a size of 112 bp). T2, target gene 2 (Human-MMP9) was positive in the MDA-MB-231 cells with a size of 139 bp). T3, target gene 3 (Mus-MMP2) was positive in the metaphysis of the tibiae of BALB/c nu/nu mice with a size of 140 bp). T4, target gene 4 (Mus-MMP9 was positive in the diaphysis of the tibiae of BALB/c nu/nu mice with a size of 229 bp. MMP, matrix metalloproteinase.
Mentions: The present study performed RT-PCR to investigate the source of MMP2 and MMP9. The results revealed that MMP2 and MMP9 mRNA were expressed in the MDA-MB-231 cells, metaphysis and diaphysis of the tibiae of the BALB/c nu/nu mice without tumor cell administration (Fig. 4).

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