Quantification of matrix metalloproteinases and tissue inhibitors of metalloproteinase in prostatic tissue: Analytical aspects

✍ Scribed by Jung, Klaus; Lein, Michael; Ulbrich, Norbert; Rudolph, Birgit; Henke, Wolfgang; Schnorr, Dietmar; Loening, Stefan A.

• Publisher: John Wiley and Sons
• Year: 1998
• Tongue: English
• Weight: 88 KB
• Volume: 34
• Category: Article
• ISSN: 0270-4137
• DOI: 10.1002/(sici)1097-0045(19980201)34:2<130::aid-pros8>3.0.co;2-o

✦ Synopsis

Background:

The balance between matrix metalloproteinases (mmp) and the tissue inhibitors of metalloproteinases (timp) has been seen as important during tumor invasion and progression. the determination of these components needs a special strategy of tissue preparation. this analytical problem has not been considered for prostatic tissue.

Methods:

We adapted an extraction method consisting of two extraction steps with 0.25% triton x-100/cacl2 solution and two heat extraction steps at 60 degrees c for 4 min. this combination allowed a complete extraction of mmp (measured as enzyme activity) and timp-1 (measured with an elisa test) from cancerous and normal prostatic tissue samples.

Results:

The median values for cancerous vs. normal mmps (50.8 mu/g wet tissue and 1,580 mu/g protein vs. 88.8 and 2,497) and timp-1 (4.49 micrograms/g wet tissue and 96.7 micrograms/g protein vs. 12.4 and 237.8) were significantly lower, whereas the respective ratios for mmp/ timp-1 (11.1 vs. 4.0 on wet weight and 15.5 vs. 5.3 on protein basis) were significantly higher.

Conclusions:

An optimized extraction procedure was elaborated for determining mmps and timp-1 in prostatic tissue samples. the increased ratio of mmp/timp-1 can be interpreted as an indicator of the imbalance between mmp and timp, characteristic of prostate carcinoma tissue.