目的明确叉头框C2(FOXC2)在结直肠癌细胞侵袭迁移中的作用。方法采用逆转录病毒感染的方法建立FOXC2稳定过表达的结直肠癌细胞株(SW480/FOXC2)及空载体对照细胞株(SW480/pBabe),显微镜下观察结直肠癌细胞形态改变。Western blot及免疫荧光法检测FOXC2过表达细胞株及对照细胞株中E-cadherin、Vimentin、N-cadherin的表达情况;Transwell侵袭小室实验检测结直肠癌细胞迁移能力的改变。结果过表达后SW480细胞的形态发生了明显的变化,从原来典型的上皮细胞形状变成长梭形,类似于成纤维细胞的形态;Western Blot及免疫荧光检测结果显示,FOXC2过表达后上皮分子标志物E-cadherin表达明显下调,而间质分子标志物Vimentin及N-cadherin表达显著上调;Transwell侵袭实验结果显示,FOXC2过表达后结直肠癌细胞侵袭潜能明显增强。结论 FOXC2过表达能诱导结直肠癌细胞SW480发生上皮-间质转化并增强其侵袭能力。
Objective To identify the role of FOXC2 in the invasion and migration of colorectal cancer cells.Methods Stable cell lines expressing FOXC2(SW480/FOXC2)or vector(SW480/pBabe)were established using retroviral infection method.The morphology alterations of SW480 cells were observed using a microscope.Western blot analysis and immunofluorescence staining assays were performed to detect the expression of E-cadherin,Vimentin and N-cadherin.The invasive and migratory abilities of colorectal cancer cells evaluated using Transwell invasion chamber experiment detection.Results The morphology of SW480 cells was significantly changed after overexpression.From the original shape typical of epithelial cells became spindle shaped growth,similar to the morphology of fibroblasts.Western blot analysis and immunofluorescence staining displayed that overexpression of FOXC2 led to significant downregulation of the epithelial marker E-cadherin,but upregulation of the mesenchymal markers Vimentin and N-cadherin.Transwell assay reveals that overexpression of FOXC2 strongly enhanced the migratory and invasive ability of SW480 cells.Conclusion FOXC2 induces epithelial-mesenchymal transition and promotes the invasive ability of colorectal cancer cells.