Establishment of a highly metastatic human lung cancer cell line and investigaion of mechanism associating with the highly metastatic ability

整合素是细胞与细胞外基质黏附的主要受体，在肿瘤转移中发挥重要作用。为了探讨整合素与肿瘤转移的关系，我们通过反复静脉注射亲本细胞(PC9)，建立了裸鼠高转移人非小细胞肺癌(NSCLC)细胞系，命名为PC9/F9。在细胞黏附实验中，PC9黏附于层粘连蛋白，而PC9/F9黏附更强。此外，PC9/F9还与IV型胶原和纤维连接蛋白黏附。流式细胞仪扫描显示VLA-2、VLA-3和VLA-6在PC9中表达，VLA-2、VLA-3、VLA-4、VLA-5和VLA-6在PC9/F9中表达。在黏附抑制实验中，抗人α3和β1整合素单抗可抑制PC9与层粘连蛋白的黏附，而抗β1整合素单抗和任何抗α整合素单抗均不抑制PC9/F9与层粘连蛋白的黏附。抗α2和抗β1整合素Mo-Ab可抑制PC9/F9与IV型胶原的黏附。抗α5和抗β1整合素Mo-Ab可抑制PC9/F9与纤维连接蛋白的黏附。抗α4Mo-Ab可适度抑制PC9/F9与纤维连接蛋白的黏附。这些数据表明，β1整合素通过改变人NSCLC细胞的表达和功能来增强其转移能力。

<ABSTRACT>Integrins are the major receptors by which cells attach to extracellular matrix and play an important role in cancer metastasis. To investigate the relation of integrins and metastasis, we established a highly metastatic human non-small cell lung cancer (NSCLC) cell line in nude mice, designated PC9/F9, by repeated intravenous injection of parent cell (PC9). In cell adhesion assay, PC9 adhered to laminin but PC9/F9 adhered more strongly. Farther more, PC9/F9 adhered type IV collagen and fibronectin. FACS scan analysis showed expression of VLA-2, VLA-3 and VLA-6 in PC9, VLA-2, VLA-3, VLA-4, VLA-5 and VLA-6 in PC9/F9. In adhesion inhibition assay, adhesion of PC9 to laminin was inhibit by anti-human alpha3 and beta1 integrin monoclonal antibody (Mo-Ab), on the other hand, adhesion of PC9/F9 to laminin was not inhibit by anti-beta1 integrin Mo-Ab and any of anti-alpha integrin Mo-Ab. Adhesion of PC9/F9 to type IV collagen was inhibited by anti-alpha2 and anti-beta1 integrin Mo-Ab. Adhesion of PC9/F9 to fibronectin was inhibit by anti-alpha5 and anti-beta1 integrin Mo-Ab. Anti-alpha4 Mo-Ab moderately inhibit adhesion of PC9/F9 to fibronectin. These data suggest that beta1 integrins enhance the ability of human NSCLC cells to metastasize by changing their expression and function.