目的:研究IQGAP1基因干扰对人食管癌细胞同质粘附能力的影响。方法:体外培养人食管癌KYSE150和EC9706细胞,利用Western blot方法检测两株细胞。IQGAP1蛋白的表达,利用缓慢聚集和细胞分离实验比较两株细胞同质粘附能力的差异;进一步在KYSE150和EC9706细胞中构建IQGAP1基因干扰的稳定细胞系,观察IQGAP1基因干扰后细胞同质粘附能力的改变。结果:KYSE150细胞IQGAP1蛋白表达量低于EC9706细胞,而同质粘附能力高于EC9706细胞;IQGAP1基因干扰后,其蛋白表达量明显降低,而细胞同质粘附能力明显增强。结论:IQGAP1基因干扰能够显著增强食管癌细胞的同质粘附能力,从而降低肿瘤细胞的恶性表型。
Objective : investigate homotypic cell-cell adhesion of esophageal detect the protein dissociation assay expression of IQGAP1 were performed to detect the effect of IQGAP1 knockdown by RNA interference (RNAi) on carcinoma (EC) cells. Method: Western blot analysis was adopted to in KYSE150 and EC9706 cells. Slow aggregation assay and cell the homotypic cell-cell adhesion properties of tumor cells. Further,KYSE150 and EC9706 cells were transfected with RNAi plasmid for targeting IQGAP1 gene or negative control, and then the cell lines of IQGAP1 knockdown were screened with G418. Level of IQGAP1 protein in interference group and control group was measured by Western blot, and the changes of homotypic cell-cell adhesion were observed by slow aggregation assay and cell dissociation assay. Result: Compared with EC9706 cells, level of IQGAP1 protein in KYSEI50 cells was lower, while the homotypic cell-cell adhesion in KYSE150 cells was stronger. Further, Western blotting analysis showed that IQGAP1 protein expression was efficiently downregulated by stable transfection with IQGAP1 siRNA expression vector. The homotypic cell-cell adhesion was obviously enhanced by IQGAP1 knockdown. Conclusion: These data suggest that downregulation of IQGAP1 expression could significantly enhance homotypic cell-cell adhesion and decrease malignant phenotype of EC cells.