Leight Jennifer L, Tokuda Emi Y, Jones Caitlin E, Lin Austin J, Anseth Kristi S
Department of Chemical and Biological Engineering, BioFrontiers Institute, and Howard Hughes Medical Institute, University of Colorado Boulder, Boulder, CO 80309; and Department of Biomedical Engineering and Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210
Department of Chemical and Biological Engineering, BioFrontiers Institute, and.
Proc Natl Acad Sci U S A. 2015 Apr 28;112(17):5366-71. doi: 10.1073/pnas.1505662112. Epub 2015 Apr 13.
Matrix metalloproteinases (MMPs) are important for many different types of cancer-related processes, including metastasis. Understanding the functional impact of changes in MMP activity during cancer treatment is an important facet not typically evaluated as part of preclinical research. With MMP activity being a critical component of the metastatic cascade, we designed a 3D hydrogel system to probe whether pharmacological inhibition affected human melanoma cell proteolytic activity; metastatic melanoma is a highly aggressive and drug-resistant form of skin cancer. The relationship between MMP activity and drug treatment is unknown, and therefore we used an in situ fluorogenic MMP sensor peptide to determine how drug treatment affects melanoma cell MMP activity in three dimensions. We encapsulated melanoma cells from varying stages of progression within PEG-based hydrogels to examine the relationship between drug treatment and MMP activity. From these results, a metastatic melanoma cell line (A375) and two inhibitors that inhibit RAF (PLX4032 and sorafenib) were studied further to determine whether changes in MMP activity led to a functional change in cell behavior. A375 cells exhibited increased MMP activity despite an overall decrease in metabolic activity with PLX4032 treatment. The changes in proteolytic activity correlated with increased cell elongation and increased single-cell migration. In contrast, sorafenib did not alter MMP activity or cell motility, showing that the changes induced by PLX4032 were not a universal response to small-molecule inhibition. Therefore, we argue the importance of studying MMP activity with drug treatment and its possible implications for unwanted side effects.
基质金属蛋白酶（MMPs）在许多不同类型的癌症相关过程中都很重要，包括转移。了解癌症治疗期间MMP活性变化的功能影响是临床前研究通常不会评估的一个重要方面。由于MMP活性是转移级联反应的关键组成部分，我们设计了一种三维水凝胶系统，以探究药物抑制是否会影响人黑色素瘤细胞的蛋白水解活性；转移性黑色素瘤是一种极具侵袭性和耐药性的皮肤癌形式。MMP活性与药物治疗之间的关系尚不清楚，因此我们使用一种原位荧光MMP传感器肽来确定药物治疗如何在三维空间中影响黑色素瘤细胞的MMP活性。我们将处于不同进展阶段的黑色素瘤细胞封装在基于聚乙二醇的水凝胶中，以研究药物治疗与MMP活性之间的关系。根据这些结果，我们进一步研究了一种转移性黑色素瘤细胞系（A375）和两种抑制RAF的抑制剂（PLX4032和索拉非尼），以确定MMP活性的变化是否导致细胞行为的功能改变。尽管用PLX4032治疗后代谢活性总体下降，但A375细胞的MMP活性却增加了。蛋白水解活性的变化与细胞伸长增加和单细胞迁移增加相关。相比之下，索拉非尼并未改变MMP活性或细胞运动性，这表明PLX4032诱导的变化并非对小分子抑制的普遍反应。因此，我们强调了研究药物治疗时MMP活性及其对不良副作用可能影响的重要性。
