Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av, Prof, Lineu Prestes 1524, Ed Biomédicas 1 sala 428, São Paulo, SP, 05508-000, Brazil.
Mol Cancer. 2013 Jan 5;12:2. doi: 10.1186/1476-4598-12-2.
ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motifs) is a member of the ADAMTS family of metalloproteases. Here, we investigated mRNA and protein levels of ADAMTS-1 in normal and neoplastic tissues using qPCR, immunohistochemistry and immunoblot analyses, and we addressed the role of ADAMTS-1 in regulating migration, invasion and invadopodia formation in breast tumor cell lines.
In a series of primary breast tumors, we observed variable levels of ADAMTS-1 mRNA expression but lower levels of ADAMTS-1 protein expression in human breast cancers as compared to normal tissue, with a striking decrease observed in high-malignancy cases (triple-negative for estrogen, progesterone and Her-2). This result prompted us to analyze the effect of ADAMTS-1 knockdown in breast cancer cells in vitro. MDA-MB-231 cells with depleted ADAMTS-1 expression demonstrated increased migration, invasion and invadopodia formation. The regulatory mechanisms underlying the effects of ADAMTS-1 may be related to VEGF, a growth factor involved in migration and invasion. MDA-MB-231 cells with depleted ADAMTS-1 showed increased VEGF concentrations in conditioned medium capable of inducing human endothelial cells (HUVEC) tubulogenesis. Furthermore, expression of the VEGF receptor (VEGFR2) was increased in MDA-MB-231 cells as compared to MCF7 cells. To further determine the relationship between ADAMTS-1 and VEGF regulating breast cancer cells, MDA-MB-231 cells with reduced expression of ADAMTS-1 were pretreated with a function-blocking antibody against VEGF and then tested in migration and invasion assays; both were partially rescued to control levels.
ADAMTS-1 expression was decreased in human breast tumors, and ADAMTS-1 knockdown stimulated migration, invasion and invadopodia formation in breast cancer cells in vitro. Therefore, this series of experiments suggests that VEGF is involved in the effects mediated by ADAMTS-1 in breast cancer cells.
ADAMTS-1（一种带有血小板反应蛋白基序的解整合素和金属蛋白酶）是 ADAMTS 家族金属蛋白酶的成员。在这里，我们使用 qPCR、免疫组织化学和免疫印迹分析研究了正常和肿瘤组织中 ADAMTS-1 的 mRNA 和蛋白水平，并研究了 ADAMTS-1 在调节乳腺癌细胞系迁移、侵袭和侵入小体形成中的作用。
在一系列原发性乳腺癌中，我们观察到 ADAMTS-1 mRNA 表达水平存在差异，但与正常组织相比，人乳腺癌中的 ADAMTS-1 蛋白表达水平较低，在高恶性病例（雌激素、孕激素和 Her-2 三阴性）中观察到明显下降。这一结果促使我们在体外分析 ADAMTS-1 敲低对乳腺癌细胞的影响。ADAMTS-1 表达耗尽的 MDA-MB-231 细胞表现出增强的迁移、侵袭和侵入小体形成。ADAMTS-1 作用的调节机制可能与 VEGF 有关，VEGF 是一种参与迁移和侵袭的生长因子。ADAMTS-1 表达耗尽的 MDA-MB-231 细胞在条件培养基中表现出更高的 VEGF 浓度，能够诱导人内皮细胞（HUVEC）小管形成。此外，与 MCF7 细胞相比，MDA-MB-231 细胞中 VEGFR2（VEGF 受体）的表达增加。为了进一步确定 ADAMTS-1 与 VEGF 调节乳腺癌细胞之间的关系，用针对 VEGF 的功能阻断抗体预处理 ADAMTS-1 表达降低的 MDA-MB-231 细胞，然后在迁移和侵袭测定中进行测试；两者均部分恢复至对照水平。
ADAMTS-1 在人乳腺癌肿瘤中表达降低，ADAMTS-1 敲低刺激乳腺癌细胞体外迁移、侵袭和侵入小体形成。因此，这一系列实验表明 VEGF 参与了 ADAMTS-1 在乳腺癌细胞中介导的作用。
