CRKL promotes lung cancer cell invasion through ERK-MMP9 pathway
CRKL promotes lung cancer cell invasion through ERK-MMP9 pathway
复制标题
CRKL通过ERK-MMP9通路促进肺癌细胞侵袭
DOI：
10.1002/mc.22148
复制
发表时间：
2015-06-01
影响因子：
4.6
通讯作者：
Wang Yan
中科院分区：
文献类型：
--
作者：
Fu Lin;Xie Chengyao;Wang Yan
CRKL是近年来发现的一种新的癌基因，在肺癌的发生发展中起重要作用。然而，CRKL在人非小细胞肺癌细胞侵袭中的潜在机制尚不清楚。我们采用免疫组化、质粒转染、Western blotting、实时荧光定量PCR、基质胶侵袭实验、染色质免疫沉淀实验和荧光素酶报告基因实验研究了CRKL在肺癌细胞侵袭中的潜在机制。CRKL在淋巴结转移癌中的表达高于原发癌。CRKL过表达增强HBE和H1299细胞系中的细胞侵袭和MMP 9表达。CRKL过表达与MMP 9高表达在原发肿瘤中有显著相关性。MMP-9抗体处理显著阻断细胞侵袭。CRKL过表达还激活AP-1荧光素酶报告活性、ERK磷酸化和c-fos与MMP 9启动子的结合。在CRKL转染的细胞中用ERK抑制剂PD 98059处理抑制ERK活性、细胞侵袭和MMP 9表达。这些结果表明CRKL过表达通过上调MMP 9表达和激活ERK通路促进细胞侵袭。(c)2014 Wiley Periodicals，Inc.
CRKL is recently defined as a new oncogene, which plays a role in the lung cancer progression. However, the potential mechanism of CRKL in human non-small cell lung cancer cell invasion is obscure. We investigated the potential mechanism of CRKL in lung cancer cell invasion using immunohistochemistry, plasmid transfection, Western blotting, real-time PCR, matrigel invasion assay, chromatin immunoprecipitation assay, and luciferase reporter assay. CRKL expression is higher in lymph node metastatic tumor compared with primary tumor. CRKL overexpression enhanced cell invasion and MMP9 expression in both HBE and H1299 cell lines. There was a significant correlation between CRKL overexpression and high MMP9 expression in primary tumors. MMP-9 antibody treatment significantly blocked cell invasion. CRKL overexpression also activated AP-1 luciferase reporter activity, ERK phosphorylation and association of c-fos to MMP9 promoter. Treatment with ERK inhibitor PD98059 in cells with CRKL transfection inhibited ERK activity, cell invasion, and MMP9 expression. These results suggested that overexpression of CRKL promoted cell invasion through upregulation of MMP9 expression and activation of ERK pathway. (c) 2014 Wiley Periodicals, Inc.