Xu Chun-Yan, Liu Shi-Quan, Qin Meng-Bin, Zhuge Chun-Feng, Qin Lin, Qin Nan, Lai Ming-Yu, Huang Jie-An
Department of Gastroenterology, Τhe First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.
Int J Mol Med. 2017 May;39(5):1277-1284. doi: 10.3892/ijmm.2017.2921. Epub 2017 Mar 17.
Sphingosine kinase 1 (SphK1) plays an important role in colorectal carcinoma metastasis. However, whether SphK1 modulates epithelial-mesenchymal transition (EMT)-related marker expression and the underlying mechanisms remain unclear. In this study, in order to clarify this issue, we used various colorectal cancer (CRC) cell lines, Caco2, HT29, RKO and HCT116. Each of the cell lines was divided into 3 groups as follows: the control group, SKI-Ⅱ (SphK1 inhibitor) group and PF-562271 [focal adhesion kinase (FAK) inhibitor] group. The migratory ability of the cells was examined by Transwell chamber assay. The mRNA and protein expression levels of SphK1, FAK (p-FAK), Slug, vimentin, N-cadherin and E-cadherin were detected by PCR and western blot analysis, respectively. The results revealed that the suppression of SphK1 reduced the cell migratory ability, and decreased the expression of Slug, vimentin and N-cadherin; however, the expression of E-cadherin was increased. Moreover, the inhibition of SphK1 reduced the expression of p-FAK. The inhibition of FAK (p-FAK) also decreased the cell migratory ability, and decreased the expression of Slug, vimentin and N-cadherin, whereas the expression of E-cadherin was increased. Thus, our data suggest that SphK1 modulates the expression of EMT-related markers and cell migration by regulating the expression of p-FAK in CRC cells. Thus, SphK1 may play a functional role in mediating the EMT process in CRC.
鞘氨醇激酶1（SphK1）在结直肠癌转移中起重要作用。然而，SphK1是否调节上皮-间质转化（EMT）相关标志物的表达及其潜在机制仍不清楚。在本研究中，为了阐明这个问题，我们使用了各种结直肠癌（CRC）细胞系，Caco2、HT29、RKO和HCT116。每个细胞系分为以下3组：对照组、SKI-Ⅱ（SphK1抑制剂）组和PF-562271[粘着斑激酶（FAK）抑制剂]组。通过Transwell小室试验检测细胞的迁移能力。分别通过PCR和蛋白质印迹分析检测SphK1、FAK（p-FAK）、Slug、波形蛋白、N-钙黏蛋白和E-钙黏蛋白的mRNA和蛋白质表达水平。结果显示，抑制SphK1可降低细胞迁移能力，并降低Slug、波形蛋白和N-钙黏蛋白的表达；然而，E-钙黏蛋白的表达增加。此外，抑制SphK1可降低p-FAK的表达。抑制FAK（p-FAK）也可降低细胞迁移能力，并降低Slug、波形蛋白和N-钙黏蛋白的表达，而E-钙黏蛋白的表达增加。因此，我们的数据表明，SphK1通过调节CRC细胞中p-FAK的表达来调节EMT相关标志物的表达和细胞迁移。因此，SphK1可能在介导CRC的EMT过程中发挥功能性作用。
