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Updated: Jun 25, 2026
Measuring Global Cellular Matrix Metalloproteinase and Metabolic Activity in 3D Hydrogels
Published on: January 22, 2019
Inhibition of hyaluronan synthases decreases matrix metalloproteinase-7 (MMP-7) expression and activity
Kelli M Bullard Dunn1, Peter K Lee, Christopher M Wilson
1Department of Surgery and Surgical Oncology, Roswell Park Cancer Institute, University at Buffalo/State University of New York, Buffalo, NY 14263, USA. kelli.bullarddunn@roswellpark.org
Introduction:
Hyaluronan (HA) and its biosynthetic enzymes hyaluronan synthases (HAS2 and HAS3) mediate Matrigel invasion by SW620 colon carcinoma cells. Because matrix metalloproteinases (MMPs) have been implicated in cancer invasion, we hypothesized that changes in HAS expression would alter MMP expression and activity in these cells.
Methods:
To determine whether an MMP was involved in invasion, Matrigel invasion assays with SW620 cells were performed in the presence or absence of the MMP inhibitors GM6001 or TIMP2. HAS isozymes were inhibited by stably transfecting SW620 cells with vectors that contained antisense HAS2 and/or -3 cDNA; transfection with an empty vector served as a control. MMP-7 transcription was assessed by quantitative reverse transcription polymerase chain reaction (RT-PCR). MMP-7 protein was detected by enzyme-linked immunosorbent assay (ELISA) and enzymatic activity compared using zymography.
Results:
GM6001 and TIMP2 decreased Matrigel invasion, which confirms that an MMP played a key role in this process. MMP-7 expression was then detected in SW620 cells. Finally, MMP-7 expression, protein, and enzymatic activity were significantly lower in antisense HAS tranfectants than in SW620 or vector control cells.
Conclusion:
We have demonstrated previously that inhibition of HAS expression and HA production in SW620 colon carcinoma cells inhibits Matrigel invasion. In the studies presented here, we have demonstrated that SW620 cells express high levels of MMP-7 and that inhibition of HAS isozymes dramatically decreases MMP-7 expression, protein, and enzymatic activity. Taken together, these findings suggest that HAS and HA may mediate cellular invasion via changes in MMP-7 expression.
Insights
Hyaluronan synthases (HAS) regulate colon cancer cell invasion by affecting matrix metalloproteinase-7 (MMP-7). Inhibiting HAS reduces MMP-7 expression and activity, suggesting a role for HAS in cancer cell invasion.
Area of Science:
- Cancer Biology
- Molecular Oncology
- Biochemistry
Background:
- Hyaluronan (HA) and its enzymes (HAS2, HAS3) are crucial for SW620 colon carcinoma cell invasion.
- Matrix metalloproteinases (MMPs) are implicated in cancer invasion, prompting investigation into their link with HAS.
Purpose of the Study:
- To investigate the hypothesis that altered hyaluronan synthase (HAS) expression impacts matrix metalloproteinase (MMP) expression and activity in SW620 colon carcinoma cells.
- To determine the role of MMPs in Matrigel invasion mediated by SW620 cells.
Main Methods:
- Matrigel invasion assays were conducted with MMP inhibitors (GM6001, TIMP2).
- Stable transfection of SW620 cells with antisense HAS2/HAS3 cDNA was used to inhibit HAS isozymes.
- MMP-7 transcription, protein levels, and enzymatic activity were quantified using RT-PCR, ELISA, and zymography, respectively.
Main Results:
- MMP inhibition significantly reduced SW620 cell Matrigel invasion.
- SW620 cells were found to express MMP-7.
- Inhibition of HAS isozymes led to a significant decrease in MMP-7 expression, protein levels, and enzymatic activity.
Conclusions:
- SW620 colon carcinoma cells express high levels of MMP-7.
- Inhibition of HAS expression and subsequent HA production dramatically reduces MMP-7 expression and activity.
- These findings suggest that HAS and HA mediate cellular invasion through modulation of MMP-7 expression.
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