Galectin-3 cleavage: A novel surrogate marker for matrix metalloproteinase activity in growing breast cancers
Galectin-3 cleavage: A novel surrogate marker for matrix metalloproteinase activity in growing breast cancers
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DOI：
10.1158/0008-5472.can-07-3233
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发表时间：
2007-12-15
期刊：
影响因子：
11.2
通讯作者：
Raz, Avraham
中科院分区：
文献类型：
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作者：
Nangia-Makker, Pratima;Raz, Tirza;Raz, Avraham
针对癌症患者基质金属蛋白酶（MMP）的治疗失败，部分原因可能是缺乏区分前基质金属蛋白酶和活性基质金属蛋白酶的诊断工具，这表明治疗是否有效。由于半乳糖凝集素-3在体外可被MMPs切割，我们开发了识别其切割和非切割形式的差异抗体，并测试了它们作为生长中的乳腺癌中活性MMPs存在的替代诊断标记物的临床应用。构建了野生型和抗裂型半乳糖凝集素-3，并在半乳糖凝集素-3缺失的人乳腺癌细胞（BT-549）中表达。通过注射裸鼠，研究了克隆的致瘤性和血管生成潜能。通过免疫组织化学方法对石蜡包埋切片进行特异性抗体分析，确定了MMP-2、MMP-9、全长和裂解的半乳糖凝集素-3在异种移植物中的位置。MMP-2/9的活性通过原位酶谱法在冷冻组织切片上得到证实。在小鼠异种移植物和人类乳腺癌标本中，半乳糖凝集素-3在体内通过差异抗体染色显示了切割，并与预测的活性MMPs共定位。原位酶谱法证实了这些结果。此外，与野生型相比，含有不可切割半凝集素-3的BT-549细胞显示出肿瘤生长和血管生成的减少。我们得出结论，半乳糖凝集素-3的裂解在肿瘤进展过程中是一个活跃的过程，可以作为生长中的乳腺癌中MMPs活性的简单、快速和可靠的替代标记物。
Failed therapies directed against matrix metalloproteinases (MMP) in cancer patients may be attributed, in part, to lack of diagnostic tools to differentiate between pro-MMPs and active MMPs, which indicate whether a treatment is efficacious or not. Because galectin-3 is cleavable in vitro by MMPs, we have developed differential antibodies recognizing its cleaved and noncleaved forms and tested their clinical utilization as a surrogate diagnostic marker for the presence of active MMPs in growing breast cancers. Wild-type and cleavage-resistant galectin-3 were constructed and expressed in galectin-3-null human breast carcinoma cells (BT-549). Tumorigenic and angiogenic potential of the clones was studied by injections into nude mice. MMP-2, MMP-9, full-length, and cleaved galectin-3 were localized in the xenografts by immunohistochemical analysis of paraffin-embedded sections using specific antibodies. Activities of MMP-2/9 were corroborated by in situ zymography on frozen tissue sections. Galectin-3 cleavage was shown in vivo by differential antibody staining and colocalized with predicted active MMPs both in mouse xenografts and human breast cancer specimens. In situ zymography validated these results. In addition, BT-549 cells harboring noncleavable galectin-3 showed reduced tumor growth and angiogenesis compared with the wild-type. We conclude that galectin-3 cleavage is an active process during tumor progression and could be used as a simple, rapid, and reliable surrogate marker for the activities of MMPs in growing breast cancers.