Regulation of matrix metalloproteinases (MMPs) expression and secretion in MDA-MB-231 breast cancer cells by LIM and SH3 protein 1 (LASP1).
Regulation of matrix metalloproteinases (MMPs) expression and secretion in MDA-MB-231 breast cancer cells by LIM and SH3 protein 1 (LASP1).
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DOI：
10.18632/oncotarget.11720
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发表时间：
2016-09-27
期刊：
影响因子：
--
通讯作者：
Butt E
中科院分区：
文献类型：
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作者：
Endres M;Kneitz S;Orth MF;Perera RK;Zernecke A;Butt E
肿瘤侵袭的过程需要蛋白水解酶降解细胞外基质。癌细胞形成突出的侵袭伪足，产生并释放基质金属蛋白酶（MMP）来降解基底膜，从而实现转移。我们研究了 LASP1（一种新发现的侵袭伪足蛋白）对侵袭性乳腺肿瘤细胞系中 MMP 表达、分泌和激活的影响。通过分析内部生成的对照和 LASP1 耗尽的 MDA-MB-231 乳腺癌细胞的微阵列数据，我们观察到 LASP1 耗尽后 MMP1、-3 和 -9 的下调。这通过蛋白质印迹分析得到证实。相反，救援实验部分恢复了 MMP 的表达和分泌。在 BT-20 乳腺癌细胞以及前列腺癌细胞系和膀胱癌细胞系中也观察到了 LASP1 对 MMP 表达的调节作用。根据我们对数据的生物信息学 FunRich 分析（绘制了 LASP1 对转录因子的高度调节），公共微阵列数据分析检测到 LASP1 高表达与 c-Fos 水平增强之间的相关性，c-Fos 是转录因子 AP-1 的一部分，已知可调节 MMP 表达。同样，在荧光素酶报告基因检测中，LASP1 敲低后 AP-1 显示转录活性降低。酶谱分析和蛋白质印迹分析显示，LASP1 进一步促进 MMP 分泌到细胞外基质中，因此很可能改变癌症进展过程中的微环境。新发现的 LASP1 通过影响 MMP 转录和分泌来调节基质降解的作用，阐明了早期研究中 LASP1 过表达侵袭性肿瘤细胞的迁移潜力。
The process of tumor invasion requires degradation of extracellular matrix by proteolytic enzymes. Cancer cells form protrusive invadopodia, which produce and release matrix metalloproteinases (MMPs) to degrade the basement membrane thereby enabling metastasis. We investigated the effect of LASP1, a newly identified protein in invadopodia, on expression, secretion and activation of MMPs in invasive breast tumor cell lines. By analyzing microarray data of in-house generated control and LASP1-depleted MDA-MB-231 breast cancer cells, we observed downregulation of MMP1, -3 and -9 upon LASP1 depletion. This was confirmed by Western blot analysis. Conversely, rescue experiments restored in part MMP expression and secretion. The regulatory effect of LASP1 on MMP expression was also observed in BT-20 breast cancer cells as well as in prostate and bladder cancer cell lines. In line with bioinformatic FunRich analysis of our data, which mapped a high regulation of transcription factors by LASP1, public microarray data analysis detected a correlation between high LASP1 expression and enhanced c-Fos levels, a protein that is part of the transcription factor AP-1 and known to regulate MMP expression. Compatibly, in luciferase reporter assays, AP-1 showed a decreased transcriptional activity after LASP1 knockdown. Zymography assays and Western blot analysis revealed an additional promotion of MMP secretion into the extracellular matrix by LASP1, thus, most likely, altering the microenvironment during cancer progression. The newly identified role of LASP1 in regulating matrix degradation by affecting MMP transcription and secretion elucidated the migratory potential of LASP1 overexpressing aggressive tumor cells in earlier studies.