FOXC1通过Rap1信号通路介导结肠癌细胞增殖与凋亡的机制研究

FOXC1 mediates the proliferation and apoptosis of colon cancer cells through the Rap1 signaling pathway

    摘要
  • 摘要:
    目的 探讨FOXC1在结肠癌中的表达特征、临床意义及其调控细胞增殖与凋亡的分子机制。
    方法 采用GEPIA数据库分析FOXC1在结肠癌中的表达及预后相关性；qRT-PCR和Western blot检测FOXC1在结肠癌细胞（HCT116、SW620）和正常结肠上皮细胞（NCM460）中的差异表达，并构建FOXC1敲低（sh-FOXC1）稳转细胞系。应用Western blot、流式细胞术、CCK-8及平板克隆实验，分析FOXC1敲低对细胞增殖、周期及凋亡的影响；通过Rap1过表达回复实验验证其下游信号通路机制。
    结果 FOXC1在结肠癌组织中的mRNA表达显著高于正常组织（P<0.001），FOXC1过表达与肿瘤分期的相关性接近显著（P=0.053），且高表达患者总体生存期缩短（Log-rank P=0.013）。敲低FOXC1后，Cyclin D1、Bcl-2表达降低，Bax表达升高（P<0.01），G0/G1期比例增加，S期减少（P<0.001），细胞增殖活性及集落形成数目减少（P<0.001）。机制研究表明，FOXC1敲低后，Rap1表达降低，Rap1GAP表达升高（P<0.05）。敲低FOXC1又恢复Rap1表达后可逆转Cyclin D1、Bcl-2的表达下调及Bax表达升高（P<0.05），S期比例增加（P<0.05），细胞增殖活性和集落形成能力增强。
    结论 FOXC1通过促进Rap1表达并下调Rap1GAP，促进结肠癌进展，靶向干预FOXC1-Rap1信号轴可能成为潜在治疗策略。

     

    Abstract:
    Objective To investigate the expression characteristics and clinical significance of FOXC1 in colon cancer, and decipher its molecular mechanism in regulating tumor cell proliferation and apoptosis.
    Methods The GEPIA database was employed to analyze the expression of FOXC1 and its correlation with prognosis in colon cancer. Differential expression of FOXC1 was detected by qRT-PCR and Western blot in colon cancer cells (HCT116 and SW620) and normal colon epithelial cells (NCM460), and stable FOXC1-knockdown (sh-FOXC1) cell lines were established. Western blot, flow cytometry, CCK-8, and plate colony formation assays were performed to analyze the effects of FOXC1 knockdown on cell proliferation, cell cycle, and apoptosis. Furthermore, the downstream signaling pathway was verified using Rap1 overexpression rescue experiments.
    Results FOXC1 mRNA expression was significantly higher in colon cancer tissues than in normal tissues (P<0.001). FOXC1 overexpression was nearing significance in relation to tumor staging (P=0.053), and patients with high FOXC1 expression had a shorter overall survival (Log-rank P=0.013). After FOXC1 knockdown, the expression of CyclinD1 and Bcl-2 decreased, whereas the expression of Bax increased (P<0.01). The proportion of cells in the G0/G1 phase increased, while the proportion of cells in the S phase decreased (P<0.001), and the cell proliferation activity and number of colonies formed decreased (P<0.001). Mechanistic studies demonstrated that after FOXC1 knockdown, Rap1 expression was reduced, while the expression of Rap1GAP increased (P < 0.05). After restoration of Rap1 expression in FOXC1-knockdown cells, the downregulation of CyclinD1 and Bcl-2 expression and the increase in Bax expression were reversed (P<0.05), the S phase ratio was increased (P<0.05), and cell proliferation activity and colony formation abilities were also rescued.
    Conclusion FOXC1 promotes colon cancer progression by facilitating Rap1 expression and downregulating Rap1GAP. Targeted intervention of the FOXC1-Rap1 signaling axis may emerge as a potential therapeutic strategy.

     

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