Department of Histology and Embryology, Dalian Medical University, Dalian, China.
Biomed Pharmacother. 2012 Sep;66(6):439-47. doi: 10.1016/j.biopha.2012.03.001. Epub 2012 Mar 27.
Caveolin-1 (CAV-1), one component of caveolae, involves in multiple cellular processes and signal transductions. We previously showed that the expression of CAV-1 gene in NCI-H446 cells inhibited cell proliferation and promoted cell metastasis. Here we explore the function of CAV-1 on tumor growth and metastasis by using NCI-H460 in vitro. First, we established NCI-H460 cell line, which CAV-1 was stably knockdown. Then we investigated the effects of CAV-1 on the morphology, proliferation, cell cycle and metastasis potential for NCI-H460 cell by crystal violet stains, CCK-8, colony formation, flow cytometry, scratch-wound assay and transwell assay. Western blot was used to examine the expression changes of cyclin D1, PCNA, E-cadherin and β-catenin. Our results showed stable knockdown of CAV-1 inhibited the proliferation of NCI-H460 cells. Cell cycle of the transfected cells was arrested in G1/S phase and the expressions of cyclin D1 and PCNA protein were downregulated. Downregulation of CAV-1 promoted the migration and invasion abilities of NCI-H460 cells in vitro. The expression of β-catenin increased and the level of E-cadherin decreased. In summary, our findings provide experimental evidence that CAV-1 may function as a proproliferative and antimetastatic gene in NCI-H460 cell line.
窖蛋白-1（CAV-1）是质膜凹陷形成的囊泡（即小窝）的基本结构成分之一，参与多种细胞过程和信号转导。我们之前的研究表明，NCI-H446 细胞中 CAV-1 基因的表达抑制了细胞增殖并促进了细胞转移。在这里，我们通过体外实验研究了 CAV-1 对 NCI-H460 细胞肿瘤生长和转移的作用。首先，我们建立了 CAV-1 稳定敲低的 NCI-H460 细胞系。然后，我们通过结晶紫染色、CCK-8、集落形成、流式细胞术、划痕实验和 Transwell 实验研究了 CAV-1 对 NCI-H460 细胞形态、增殖、细胞周期和转移潜能的影响。Western blot 用于检测细胞周期蛋白 D1、PCNA、E-钙粘蛋白和β-连环蛋白的表达变化。结果显示，CAV-1 的稳定敲低抑制了 NCI-H460 细胞的增殖。转染细胞的细胞周期被阻滞在 G1/S 期，细胞周期蛋白 D1 和 PCNA 蛋白的表达下调。CAV-1 的下调促进了 NCI-H460 细胞在体外的迁移和侵袭能力。β-连环蛋白的表达增加，E-钙粘蛋白的水平降低。综上所述，我们的研究结果为 CAV-1 可能在 NCI-H460 细胞系中作为促增殖和抗转移基因发挥作用提供了实验依据。
