Overview of matrix metalloproteinase expression in cultured human cells

Giambernardi, Troy A.; Grant, George M.; Taylor, Gail P.; Hay, Robert J.; Maher, Veronica M.; McCormick, J. Justin; Klebe, Robert J.

doi:10.1016/s0945-053x(98)90019-1

Cited by 242 publications

(192 citation statements)

References 60 publications

Supporting

Mentioning

179

Contrasting

Unclassified

Order By: Relevance

“…Previous studies showed that the PC-3 cell line expresses uPA (Hoosein et al, 1991) and mRNA for 10 MMPs including MMP-1 and MMP-7 (Giambernardi et al, 1998). We confirmed that the PC-3 cell line gave strong positive staining for uPA, MMP-1 and MMP-7.…”

Section: Discussionsupporting

confidence: 84%

Role of proteolytic enzymes in human prostate bone metastasis formation: in vivo and in vitro studies

et al. 2002

View full text Add to dashboard Cite

Prostate cancers ability to invade and grow in bone marrow stroma is thought to be due in part to degradative enzymes. The formation of prostate skeletal metastases have been reproduced in vitro by growing co-cultures of prostatic epithelial cells in bone marrow stroma. Expression of urokinase plasminogen activator, matrix metalloproteinase 1 and 7 by prostatic epithelial cells were identified using immunocytochemistry. Also, in vivo tissue sections from human prostatic bone marrow metastases were stained. To establish the role of these enzymes on colony formation, inhibitory antibodies directed against urokinase plasminogen activator, matrix metalloproteinase 1 and matrix metalloproteinase 7 were added into primary prostatic epithelial cells and bone marrow stroma co-cultures. All prostatic epithelial cell cultures stained positively for matrix metalloproteinase 1, matrix metalloproteinase 7 and urokinase plasminogen activator. Generally prostatic epithelial cells derived from malignant tissues showed increased staining in comparison to epithelia derived from non-malignant tissue. In agreement with in vitro cocultures, the in vivo tissue sections of prostate bone marrow metastases showed positive staining for all three enzymes. Inhibition studies demonstrated that blocking matrix metalloproteinase 1, matrix metalloproteinase 7 and urokinase plasminogen activator function reduced the median epithelial colony area significantly in bone marrow stroma co-cultures in vitro. Using a human ex-vivo model we have shown that matrix metalloproteinase 1, matrix metalloproteinase 7 and urokinase plasminogen activator play an important role in the establishment of prostatic epithelial cells within bone marrow.

show abstract

Section: Discussionsupporting

confidence: 84%

Role of proteolytic enzymes in human prostate bone metastasis formation: in vivo and in vitro studies

et al. 2002

View full text Add to dashboard Cite

show abstract

“…Primer sequences were from published data for MMP and TIMP amplification and are listed in Table 1. 49,50 LNCaP and PC3 cDNAs were used to optimize the conditions for RT-PCR for each primer set by comparing different concentrations of magnesium chloride and different cycle numbers. The concentration that gave the strongest signal on a 2% agarose gel stained with ethidium bromide was chosen as the optimal concentration for that primer set.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…cDNA was amplified using a HYBAID PCR machine at 94 C for 4 min (1 cycle), followed by 35 cycles of 94 C for 30 s, annealing temperature for 30 s and 72 C for 1 min, and a final step of 72 C for 10 min. 49 Amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was undertaken to monitor the quality of the RNA extraction and cDNA synthesis and to ensure that equivalent amounts of cDNA were used in MMP/ TIMP amplifications. The PCR mixture for GAPDH consisted of 5 ml of 106 polymerase buffer, 0.75 mM MgCl 2 , 0.5 mM dNTPs, 10 pmol GAPDH 5 0 primer (5 0 -CCA CCC ATG GCA AAT TCC ATG GCA-3 0 ), 10 pmol 3 0 primer (5 0 -TCT AGA CGG CAG GTC AGG TCC ACC-3 0 ), 0.5 ml Taq polymerase, 2 ml cDNA template and sterile RNase-free water to give a volume of 50 ml.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…The PCR conditions were 94 C for 4 min (1cycle), followed by 94 C for 30 s, 60 C for 30 s and 72 C for 90 s (24 cycles), and a final step of 72 C for 10 min. PCR products were separated on a 2% agarose gel stained with ethidium bromide or silver staining of a 4% acrylamide gel and quantified using a Kodak Digital Table 1 Primer sequences for MMP 49 Scientific Imaging System (Kodak, Rochester, NY, USA). Expression of MMPs relative to GADPH was estimated by densitometry.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…MMP mRNA expression was determined by RT-PCR using published primers. 49 As cell density can affect the levels of mRNA expression of some MMPs in vitro, 52 -55 we determined the optimal harvest time using PC3 cells as a representative cell line and examined expression of representative MMPs: MMP-1, MT1 and MT2-MMP by RT-PCR. PC3 cells at various cell densities were obtained by seeding increasing numbers of cells and harvesting at the same time (designated Gl -G8), or by seeding at the same density and harvesting at 24 h intervals up to 6 days (designated D1 -D6).…”

Section: In Vitro Invasion Assaysmentioning

confidence: 99%

See 2 more Smart Citations

Characterization of expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in prostate cancer cell lines

Daja

Niu

Zhao

et al. 2003

Prostate Cancer Prostatic Dis

View full text Add to dashboard Cite

Stromal expression of some matrix metalloproteinases (MMPs) has been associated with increasing tumour burden in prostate cancer. We investigated the expression of mRNA (by RT-PCR) and protein (by zymography and western blotting) of MMPs and endogenous inhibitors (tissue inhibitors of metalloproteinases, TIMPs) in two parent epithelial prostate cancer cell lines and sublines of increasing invasive/metastatic potential. Expression of membrane type MMPs, MT1-MMP and MT3-MMP mRNA was higher in PC3-derived than in LNCaPderived lines, whereas MT2-MMP mRNA expression was higher in the LNCaPderived than in PC3-derived cell lines. Active MT1, MT2 and MT3-MMP protein levels were similar in all lines, but processed MT-MMPs, indicative of latent MMP activation, were increased in more aggressive sublines. Expression of MMP-1, MMP-13 and TIMP-1 was higher in the more aggressive sublines and may be implicated in invasive/metastatic ability. Regulation of MMP-1 and MMP-13 expression may offer important therapeutic options for treating patients with prostate cancer.

show abstract

Gamma-irradiation induces matrix metalloproteinase II expression in a p53-dependent manner

Wang

Sun

2000

Mol. Carcinog.

View full text Add to dashboard Cite

Matrix metalloproteinases (MMPs) are a family of proteinases that degrade the basement membrane and have been implicated in promoting tumor metastasis. MMP-2, one member of this family, was recently found to be a p53 target and subject to p53 upregulation. In this study, we examined the correlation between the expression of MMP-2 and the increased expression of p53 after gamma-irradiation. Three human p53-positive cell lines that express wild-type p53, including U2-OS (osteosarcoma), RKO (colon carcinoma), MCF-7 (breast carcinoma), one mouse p53 positive cell line and HepG2 (liver carcinoma), and two p53-negative human cell lines, SAOS-2 (osteosarcoma) and RKO-E6 (colon carcinoma), were used in this study. The MMP-2 activity was analyzed by using gelatin zymography. The p53 level was measured by western blot analysis. Our results show that wild-type p53 induced by ionizing radiation caused a subsequent increase of MMP-2 activity in U2-OS and RKO cells but not in MCF-7, HepG2, SAOS-2, or RKO-E6 cells. These results suggest that the gamma-radiation-induced expression of MMP-2 is dependent on the cell type and presence of functional p53. Thus, ionizing radiation could activate MMP-2 activity in a subset of human cancer cells and may lead to an increase in their metastatic potential.

show abstract

Overview of matrix metalloproteinase expression in cultured human cells

Cited by 242 publications

References 60 publications

Role of proteolytic enzymes in human prostate bone metastasis formation: in vivo and in vitro studies

Role of proteolytic enzymes in human prostate bone metastasis formation: in vivo and in vitro studies

Characterization of expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in prostate cancer cell lines

Gamma-irradiation induces matrix metalloproteinase II expression in a p53-dependent manner

Contact Info

Product

Resources

About