Siqueira Adriane S, Gama-de-Souza Letícia N, Arnaud Maria Vanda C, Pinheiro João J V, Jaeger Ruy G
Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Lineu Prestes,1524 Ed. Biomédicas 1, sala 302, São Paulo, SP, 05508-000, Brazil.
Tumour Biol. 2010 Jan;31(1):46-58. doi: 10.1007/s13277-009-0008-x. Epub 2009 Dec 9.
Squamous cell carcinoma is a prevalent head and neck tumor with high mortality. We studied the role played by laminin alpha1 chain peptide AG73 on migration, invasion, and protease activity of cells (OSCC) from human oral squamous cell carcinoma. Immunohistochemistry and immunofluorescence analyzed expression of laminin alpha1 chain and MMP9 in oral squamous cells carcinoma in vivo and in vitro. Migratory activity of AG73-treated OSCC cells was investigated by monolayer wound assays and in chemotaxis chambers. AG73-induced invasion was assessed in Boyden chambers. Invasion depends on MMPs. Conditioned media from cells grown on AG73 was subjected to zymography. We searched for AG73 receptors related to these activities in OSCC cells. Immunofluorescence analyzed AG73-induced colocalization of syndecan-1 and beta1 integrin. Cells had these receptors silenced by siRNA, followed by treatment with AG73 and analysis of migration, invasion, and protease activity. Oral squamous cell carcinoma expresses laminin alpha1 chain and MMP9. OSCC cells treated with AG73 showed increased migration, invasion, and protease activity. AG73 induced colocalization of syndecan-1 and beta1 integrin. Knockdown of these receptors decreased AG73-dependent migration, invasion, and protease activity. Syndecan-1 and beta1 integrin signaling downstream of AG73 regulate migration, invasion, and MMP production by OSCC cells.
鳞状细胞癌是一种常见的头颈部肿瘤，死亡率很高。我们研究了层粘连蛋白α1链肽AG73对人口腔鳞状细胞癌细胞（OSCC）迁移、侵袭及蛋白酶活性的作用。采用免疫组织化学和免疫荧光法分析体内和体外口腔鳞状细胞癌中层粘连蛋白α1链和基质金属蛋白酶9（MMP9）的表达情况。通过单层划痕试验和趋化性小室研究AG73处理后的OSCC细胞的迁移活性。在Boyden小室中评估AG73诱导的侵袭情况。侵袭依赖于基质金属蛋白酶。对在AG73上生长的细胞的条件培养基进行酶谱分析。我们在OSCC细胞中寻找与这些活性相关的AG73受体。免疫荧光分析AG73诱导的多功能蛋白聚糖-1（syndecan-1）和β1整合素的共定位。用小干扰RNA（siRNA）使细胞中的这些受体沉默，随后用AG73处理并分析迁移、侵袭及蛋白酶活性。口腔鳞状细胞癌表达层粘连蛋白α1链和MMP9。用AG73处理的OSCC细胞显示出迁移、侵袭及蛋白酶活性增加。AG73诱导syndecan-1和β1整合素的共定位。这些受体的敲低降低了AG73依赖性迁移、侵袭及蛋白酶活性。AG73下游的syndecan-1和β1整合素信号传导调节OSCC细胞的迁移、侵袭及MMP生成。
