[Effect of Kruppel-like factor 2 on the migration of human liver sinusoidal endothelial cells].
[Effect of Kruppel-like factor 2 on the migration of human liver sinusoidal endothelial cells].
复制标题
Kruppel样因子2对人肝窦内皮细胞迁移的影响
作者：
N. Li;C. Liu;D. Pan;Y. Tseng;J. Zhou;X. Zeng;T. Luo;S. Y. Chen
目的：探讨KRUPel样因子2(KLF2)对人肝窦内皮细胞迁移的影响及其机制。方法：用不同的Lenti病毒感染体外培养的人晶状体上皮细胞，分别用不同的Lenti病毒(LV5-KLF2和LV3-shKLF2)过表达或抑制KLF2的表达，用实时荧光定量聚合酶链式反应(RT-PCR)和Western印迹法检测KLF2对LSEC迁移的影响，用实时荧光定量PCR法和Western印迹法分别检测血管内皮生长因子受体-2(VEGFR-2)的表达和蛋白表达，Western印迹法检测Src、P38MAPK和P44/42MAPK的表达和磷酸化。结果：KLF2表达上调明显抑制LSec的迁移，12h后LV5-KLF2细胞迁移至滤器下端的细胞数明显少于对照细胞[(35.6±1.4)，(71.3±2.4)，(6 9.3±1.6)，P<；0.001]。相反，KLF2的表达下调促进了LSEC的迁移，与LV3-NC和WT对照细胞相比，LV3-KLF2细胞迁移到滤器下端的细胞更多[(189.5±5)，(83 4±2 5)和(82 2±3 4)，P<；0.001]。此外，KLF2的上调降低了LSE中VEGFR2的mRNA和蛋白表达水平，而KLF2的下调显著增加了VEGFR2的表达。此外，KLF2的上调抑制了LSEC中Src、P38MAPK和p44/42MAPK通路的磷酸化，而KLF2的下调促进了这些信号通路蛋白的磷酸化。结论：KLF2可能通过Src/MAPK信号通路抑制人LSEC的迁移。
Objective: To investigate the effect and mechanism of Kruppel-like factor 2 (KLF2) on the migration of human liver sinusoidal endothelial cells (LSEC). Methods: Cultured human LSEC were infected with different lenti-viruses to overexpress or suppress KLF2 expression (LV5-KLF2 and LV3-shKLF2, respectively), the infection efficacies were examined by real-time PCR and Western blot analysis.Transwell migration assay was used to investigate the role of KLF2 on the migration of LSEC.The mRNA and protein expression of vascular endothelial growth factor receptor-2 (VEGFR-2) were detected by real-time PCR and Western blot analysis, respectively.The expression and phosphorylation of Src, P38 MAPK, and P44/42 MAPK were detected by Western blot. Results: The up-regulation of KLF2 expression dramatically inhibited migration of treated LSEC, compared with LV5-NC and WT control cells, fewer LV5-KLF2 cells migrated to the lower side of the filter after 12 h [ (35.6±1.4), (71.3±2.4) and (69.3±1.6), P<0.001 for all comparisons]. In contrast, the down-regulation of KLF2 expression promoted the migration of LSEC, more LV3-KLF2 cells migrated to the lower side of the filter compared with the LV3-NC and WT control cells [(189.5±5.4), (83.4±2.5) and (82.2±3.4), P<0.001 for all comparisons]. Furthermore, up-regulation of KLF2 reduced the mRNA and protein expression level of VEGFR2, while down-regulation of KLF2 significantly increased its expression in LSEC.Additionally, up-regulation of KLF2 inhibited the phosphorylation of Src, P38 MAPK, and P44/42 MAPK pathway in LSEC, whereas down-regulation of KLF2 promoted the phosphorylation of those signaling pathway proteins. Conclusions: KLF2 may inhibit the migration of human LSEC through the Src/ MAPK signaling pathway.