Jung K, Lein M, Ulbrich N, Rudolph B, Henke W, Schnorr D, Loening S A
Department of Urology, University Hospital Charité, Humboldt University, Berlin, Germany.
Prostate. 1998 Feb 1;34(2):130-6. doi: 10.1002/(sici)1097-0045(19980201)34:2<130::aid-pros8>3.0.co;2-o.
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.
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.
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.
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.
基质金属蛋白酶（MMP）与金属蛋白酶组织抑制剂（TIMP）之间的平衡在肿瘤侵袭和进展过程中被视为至关重要。这些成分的测定需要一种特殊的组织制备策略。对于前列腺组织，尚未考虑这一分析问题。
我们采用了一种提取方法，包括用0.25% Triton X - 100/CaCl₂溶液进行两步提取以及在60℃下进行两步4分钟的热提取。这种组合能够从癌性和正常前列腺组织样本中完全提取MMP（以酶活性测量）和TIMP - 1（用ELISA试验测量）。
癌性与正常组织的MMP中位数（湿组织50.8 mU/g和蛋白质1,580 mU/g，对比正常组织的88.8和2,497）以及TIMP - 1中位数（湿组织4.49微克/g和蛋白质96.7微克/g，对比正常组织的12.4和237.8）显著更低，而MMP/TIMP - 1的相应比率（以湿重计为11.1对比4.0，以蛋白质计为15.5对比5.3）显著更高。
精心制定了一种优化的提取程序，用于测定前列腺组织样本中的MMP和TIMP - 1。MMP/TIMP - 1比率的增加可被解释为MMP与TIMP失衡的指标，这是前列腺癌组织的特征。
