Department of Thoracic Surgery, Tangdu Hospital, Forth Military Medical University, China; Department of Cardio-thoracic Surgery, Affiliated Hospital of Academy of Military Medical Sciences, China.
Exp Cell Res. 2013 Oct 15;319(17):2545-53. doi: 10.1016/j.yexcr.2013.05.016. Epub 2013 Jun 18.
Migration-stimulating factor (MSF), an oncofetal truncated isoform of fibronectin, is a potent stimulator of cell invasion. However, its distribution and motogenic role in non-small cell lung cancer (NSCLC) have never been identified. In this study, real-time PCR and immunohistochemical staining (IHC) were performed to detect MSF mRNA and protein levels in tumor tissues and matched adjacent tumor-free tissues. Furthermore, to examine the effect of MSF on invasiveness, MSF was upregulated in A549 cells. The invasiveness and viability of A549 cells were then determined using a transwell migration assay and the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assays, respectively. The expression level of MSF in NSCLC tissue was markedly higher than in matched adjacent tumor-free tissue. Additionally, the level of MSF protein expression in stage III and IV NSCLC samples was higher than in stage I and II NSCLC samples. More importantly, we also demonstrated that migration and invasion of A549 cells increased substantially after upregulating MSF, although proliferation remained unchanged. Meanwhile, we found no correlation between increasing motility and invasiveness of MSF-overexpressing cells and expression levels and activities of matrix metalloprotease MMP-2 and MMP-9. Our current study shows that MSF plays a role in migration and invasion of A549 cells and suggests that MSF may be a potential biomarker of NSCLC progression.
迁移刺激因子 (MSF) 是纤连蛋白的一种癌胚截断异构体,是细胞侵袭的有效刺激物。然而,其在非小细胞肺癌 (NSCLC) 中的分布和促运动作用从未被确定。在这项研究中,通过实时 PCR 和免疫组织化学染色 (IHC) 检测了肿瘤组织和匹配的无肿瘤相邻组织中的 MSF mRNA 和蛋白水平。此外,为了研究 MSF 对侵袭性的影响,我们在 A549 细胞中上调了 MSF。然后使用 Transwell 迁移分析和 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐 (MTT) 活力测定分别确定 A549 细胞的侵袭性和活力。MSF 在 NSCLC 组织中的表达水平明显高于匹配的无肿瘤相邻组织。此外,III 期和 IV 期 NSCLC 样本中 MSF 蛋白表达水平高于 I 期和 II 期 NSCLC 样本。更重要的是,我们还表明,上调 MSF 后 A549 细胞的迁移和侵袭显著增加,尽管增殖保持不变。同时,我们发现 MSF 过表达细胞的运动和侵袭性增加与基质金属蛋白酶 MMP-2 和 MMP-9 的表达水平和活性之间没有相关性。我们目前的研究表明,MSF 在外源性 A549 细胞的迁移和侵袭中发挥作用,并提示 MSF 可能是 NSCLC 进展的潜在生物标志物。