CRKL plays a pivotal role in tumorigenesis of head and neck squamous cell carcinoma through the regulation of cell adhesion
CRKL plays a pivotal role in tumorigenesis of head and neck squamous cell carcinoma through the regulation of cell adhesion
复制标题
DOI：
10.1016/j.bbrc.2011.12.142
复制
发表时间：
2012-02-03
影响因子：
3.1
通讯作者：
Tanaka, Shinya
中科院分区：
文献类型：
--
作者：
Yanagi, Hiroko;Wang, Lei;Tanaka, Shinya
CRK是参与调节人类癌症恶性潜能的不可或缺的分子。CRK-like（CRKL）是CRK的一种在造血细胞中占优势的同源物，在慢性粒细胞白血病患者中被BCR-ABL酪氨酸激酶磷酸化，但其在非造血系统肿瘤中的生物学功能尚不清楚。在这项研究中，我们探讨了CRKL在头颈部鳞状细胞癌（HNSCC）的致瘤作用在体外和体内。HNSCC细胞系HSC-3细胞的免疫沉淀分析表明，C3 G的主要结合伴侣是CRKL，而不是CRK。为了阐明CRKL的分子功能，我们建立了慢病毒shRNA介导的CRKL敲低的HNSCC细胞系。在CRKL敲低的HSC-3和HSC-4细胞中，与对照细胞相比，细胞生长和运动性减弱。细胞粘附试验表明，在CRKL敲低的HSC-3细胞中，细胞粘附到纤连蛋白和胶原蛋白包被的培养皿上受到显著抑制，而聚L-赖氨酸包被的培养皿没有观察到显著变化。免疫荧光染色显示，CRKL敲低的HSC-3细胞中粘着斑减少。通过下拉试验，CRKL敲低的HSC-3细胞显示与对照细胞相比活性Rap 1的量减少。此外，在体内测定中，CRKL敲低的HSC-3细胞在裸鼠中的肿瘤形成显著消除。我们的研究结果表明，CRKL调节HNSCC细胞的生长，运动性和整合素依赖的细胞粘附，表明CRKL在HNSCC致瘤性中起主要作用。(C)2012 Elsevier Inc. All rights reserved.
The signaling adapter protein CRK is an indispensable molecule involved in regulating the malignant potential of human cancers. CRK-like (CRKL) is a hematopoietic cell-dominant homologue of CRK that is reported to be phosphorylated by BCR-ABL tyrosine kinase in chronic myelogenous leukemia patients, but its biological function in non-hematopoietic tumors remains unclear. In this study, we explored the tumorigenic role of CRKL in head and neck squamous cell carcinoma (HNSCC) in vitro and in vivo. Immunoprecipitation analysis of HNSCC cell line, HSC-3 cells, showed that the dominant binding partner for C3G was CRKL, not CRK. To clarify the molecular function of CRKL, we established lentiviral shRNA-mediated CRKL-knockdown HNSCC cell lines. In CRKL-knockdown HSC-3 and HSC-4 cells, cell growth and motility were diminished compared to control cells. Cell adhesion assays showed that cell attachment onto both fibronectin- and collagen-coated dishes was significantly suppressed in CRKL-knockdown HSC-3 cells, while no significant change was observed for poly-L-lysine-coated dishes. Immunofluorescence staining revealed that focal adhesion was reduced in CRKL-knockdown HSC-3 cells. With a pull-down assay, CRKL-knockdown HSC-3 cells showed decreased amounts of active Rap1 compared to control cells. Moreover, in an in vivo assay, tumor formation of CRKL-knockdown HSC-3 cells in nude mice was significantly abrogated. Our results indicate that CRKL regulates HNSCC-cell growth, motility, and integrin-dependent cell adhesion, suggesting that CRKL plays a principal role in HNSCC tumorigenicity. (C) 2012 Elsevier Inc. All rights reserved.