Involvement of adaptor protein Crk in malignant feature of human ovarian cancer cell line MCAS
Involvement of adaptor protein Crk in malignant feature of human ovarian cancer cell line MCAS
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DOI：
10.1038/sj.onc.1209398
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发表时间：
2006-06-15
期刊：
影响因子：
8
通讯作者：
Tanaka, S.
中科院分区：
文献类型：
--
作者：
Linghu, H.;Tsuda, M.;Tanaka, S.
信号衔接蛋白Crk通过其靶点Dock180和C3G调节细胞运动和生长，Dock180和C3G分别是小GTP酶Rac和Rap的鸟嘌呤核苷酸交换因子（GEF）。最近，已报道在各种人类癌症中Crk的过表达。敬德。由于Crk在人癌细胞中的作用，通过RNA干扰在人卵巢癌细胞系MCAS中靶向Crk表达，从而建立了三种Crk敲低细胞系。这些细胞系表现出紊乱的肌动蛋白纤维，减少了粘着斑的数量，并废除了片状伪足的形成。拉下试验和FRET为基础的延时显微镜，与抑制的运动和侵袭的吞噬动力学轨道试验和transwell试验在这些细胞中的Rac活性降低。此外，Crk敲低细胞在培养中表现出缓慢的生长速率，并抑制软琼脂中的锚定依赖性生长。当Crk敲低细胞被注射到腹腔中时，在裸鼠中的肿瘤形成潜力被减弱，并且没有观察到腹膜内扩散。这些结果表明，Crk是一个关键组成部分的粘着斑，并参与细胞的生长，侵袭和传播的人卵巢癌细胞系MCAS。
Signaling adaptor protein Crk regulates cell motility and growth through its targets Dock180 and C3G, those are the guanine-nucleotide exchange factors (GEFs) for small GTPases Rac and Rap, respectively. Recently, overexpression of Crk has been reported in various human cancers. To de. ne the role for Crk in human cancer cells, Crk expression was targeted in the human ovarian cancer cell line MCAS through RNA interference, resulting in the establishment of three Crk knockdown cell lines. These cell lines exhibited disorganized actin fibers, reduced number of focal adhesions, and abolishment of lamellipodia formation. Decreased Rac activity was demonstrated by pull-down assay and FRET-based time-lapse microscopy, in association with suppression of both motility and invasion by phagokinetic track assay and transwell assay in these cells. Furthermore, Crk knockdown cells exhibited slow growth rates in culture and suppressed anchorage-dependent growth in soft agar. Tumor forming potential in nude mice was attenuated, and intraperitoneal dissemination was not observed when Crk knockdown cells were injected into the peritoneal cavity. These results suggest that the Crk is a key component of focal adhesion and involved in cell growth, invasion, and dissemination of human ovarian cancer cell line MCAS.