Elevated expression of Cyr61 enhances peritoneal dissemination of gastric cancer cells through integrin α2β1
Elevated expression of Cyr61 enhances peritoneal dissemination of gastric cancer cells through integrin α2β1
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DOI：
10.1074/jbc.m706600200
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发表时间：
2007-11-23
影响因子：
4.8
通讯作者：
Kuo, Min-Liang
中科院分区：
文献类型：
--
作者：
Lin, Ming-Tsan;Chang, Cheng-Chi;Kuo, Min-Liang
富半胱氨酸61 （Cyr61/CCN1）参与人胃癌的发生和发展。尽管如此，Cyr61在这类癌症的腹膜播散中的作用尚未完全确定。我们利用脂质体介导转染，在胃癌AGS或MKN45细胞系中建立Cyr61或反义Cyr61表达载体。通过体外和离体癌细胞粘附试验对转染物进行了检测。此外，我们还通过功能性整合素荧光激活细胞分选实验、逆转录pcr和AP-1报告基因实验来研究Cyr61的潜在信号通路。结果表明，稳定转染Cyr61后，AGS细胞株的粘附能力明显增强。在AGS细胞内过表达Cyr61可显著提高整合素α (2) β的功能性表达(1)。整合素α (2) β(1)的功能中和抗体有效抑制cyr61介导的AGS细胞对腹膜组织的增强粘附。整合素α(2)基因启动子分析进一步表明，AP-1通路在表达cyr61的AGS细胞中被明显激活。动物研究表明，与Neo对照细胞系相比，注射cyr61过表达的AGS细胞的小鼠腹膜播种结节数量更多，存活率更低，当这些细胞被整合素α (2) β(1)的功能性阻断抗体处理时，它们能够引起腹膜播散的下降。这些数据表明Cyr61可能通过ap -1依赖性途径上调功能性整合素α (2) β(1)，从而促进肿瘤细胞粘附能力，从而促进胃癌的腹膜播散。
Cysteine-rich 61 (Cyr61/CCN1) is involved in human gastric cancer development and progression. Nonetheless, the role of Cyr61 as regards peritoneal dissemination of such cancers has not yet been completely characterized. We used liposome-mediated transfection to establish Cyr61, or antisense Cyr61, expression vectors into gastric cancer AGS or MKN45 cell lines. Transfectants were tested by means of a cancer-cell adhesion assay in vitro and ex vivo. Furthermore, a functional integrin fluorescence-activated cell sorting assay, reverse transcription-PCR, and an AP-1 reporter assay were performed to investigate the potential signaling pathway of Cyr61. It was shown that stable transfection of Cyr61 into the AGS cell line strongly enhanced its adhesion ability. The overexpression of Cyr61 within AGS cells significantly increased the functional expression of integrin alpha(2)beta(1). Function-neutralizing antibody to integrin alpha(2)beta(1) effectively suppressed the Cyr61-mediated enhanced adhesion of AGS cells to peritoneal tissue. Promoter assays of integrin alpha(2) gene further revealed that the AP-1 pathway was evidently activated within Cyr61-expressing AGS cells. Animal studies have revealed that mice injected with Cyr61-overexpressed AGS cells featured a greater number of peritoneal seeding nodules and a lower survival rate than the Neo control cell lines, and when such cells were treated with functional blocking antibody to integrin alpha(2)beta(1), they were able to elicit a decline in the peritoneal dissemination. The data suggest that Cyr61 may contribute to the peritoneal dissemination of gastric cancer by promoting tumor-cell adhesion ability through the up-regulation of the functional integrin alpha(2)beta(1) via an AP-1-dependent pathway.