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Overview of matrix metalloproteinase expression in cultured human cells
T A Giambernardi1, G M Grant, G P Taylor
1Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio 78284, USA.
Abstract:
The matrix metalloproteinases (MMP) have been implicated in tumor invasion and metastasis both by immunohistochemical studies and from the observation that specific metalloproteinase inhibitors block tumor invasion and metastasis. Oligonucleotide primers for thirteen MMPs (MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11, MMP-12, MMP-13, MMP-14, MMP-15, MMP-16) were optimized for use in RT-PCR. A semi-quantitative RT-PCR assay was used to determine the pattern of MMP mRNA expression in 84 normal and transformed or carcinogen transformed human cell lines and strains derived from different tissues. The results demonstrate one or more cell lines which express thirteen members of the MMP family. In addition, various oncogene transfected human fibroblast cell strains were analyzed for MMP expression. We confirm that over-expression of the H-ras oncoprotein correlates with up-regulation of MMP-9 and demonstrate that over-expression of v-sis also up-regulates MMP-9. A cell line immortalized following myc expression was found to up-regulate MMP-7, MMP-11 and MMP-13. Inappropriate expression of several MMP mRNAs was detected in breast, prostate, bone, colon and oral tumor derived cell lines. Identification of at least one cell line expressing each of thirteen MMPs and the observation of oncogene induced expression of several MMPs should facilitate analysis of the transcriptional mechanisms controlling each MMP.
Insights
This study optimized primers for thirteen matrix metalloproteinases (MMPs) and used RT-PCR to analyze their mRNA expression in human cell lines. Results show oncogenes upregulate MMPs, aiding future research into cancer progression.
Area of Science:
- Molecular Biology
- Cancer Research
- Biochemistry
Background:
- Matrix metalloproteinases (MMPs) are crucial in tumor invasion and metastasis.
- Inhibitors of MMPs have shown potential in blocking cancer spread.
Purpose of the Study:
- To optimize oligonucleotide primers for thirteen MMPs for RT-PCR analysis.
- To determine the mRNA expression patterns of MMPs in various human cell lines.
- To investigate the correlation between oncogene expression and MMP upregulation.
Main Methods:
- Optimization of oligonucleotide primers for thirteen MMPs (MMP-1 to MMP-16, excluding MMP-4, 5, 6, 17).
- Semi-quantitative RT-PCR assay to analyze MMP mRNA expression.
- Analysis of MMP expression in 84 normal and transformed human cell lines and oncogene-transfected fibroblasts.
Main Results:
- Successfully identified cell lines expressing all thirteen targeted MMPs.
- Confirmed that H-ras and v-sis oncoprotein overexpression upregulates MMP-9.
- Observed that myc expression upregulates MMP-7, MMP-11, and MMP-13.
- Detected aberrant expression of several MMP mRNAs in tumor-derived cell lines (breast, prostate, bone, colon, oral).
Conclusions:
- The study established a comprehensive profile of MMP mRNA expression across diverse human cell lines.
- Oncogene-driven MMP expression provides insights into transcriptional regulation mechanisms.
- Findings facilitate further investigation into the role of specific MMPs in cancer development and metastasis.