沉默CXCR4基因对食管鳞癌EC9706细胞中VEGF基因表达的影响

Effect of silencing CXCR4 on the expression of VEGF in EC9706

目的探讨沉默CXCR 4基因对食管鳞癌EC9706细胞中VEGF基因表达的影响,初步阐明CX-CR 4在食管鳞癌细胞系EC9706浸润转移中的作用机制。方法利用R T-PCR检测转染有两对CXCR 4siR NA的食管鳞癌EC9706细胞中VEGF基因mR NA的表达,Western Blot和免疫细胞化学技术检测转染有两对CXCR4 siRNA的食管鳞癌EC9706细胞中VEGF基因蛋白的表达,同时以未转染细胞和转染阴性siR-NA细胞作为空白对照和阴性对照。结果转染两对CXCR 4 siR NA后48 h,R T-PCR结果显示,转染组VEGF mR NA的表达较未转染空白对照组和转染阴性对照siR NA组相比明显下降,差异具有显著性(P<0.05);Western Blot结果显示CXCR 4 siR NA转染后48 h,CXCR 4 siR NA不仅能抑制VEGF mR NA的表达,对VEGF蛋白的表达同样有明显的抑制作用,差异具有显著性(P<0.05);免疫细胞化学技术检测结果进一步证实,与未转染空白对照组和转染阴性对照siRNA组相比,CXCR siRNA对VEGF蛋白表达具有显著抑制作用,差异有显著性(P<0.05)。结论特异性阻断CXCR4基因的表达可以抑制VEGF基因的表达,说明CX-CR 4基因有可能通过调控VEGF的表达参与食管鳞癌的浸润转移,CXCR 4有望成为肿瘤基因治疗的新靶点。

【Objective】 To explore the effect of silencing CXCR4 on the expression of VEGF in EC9706 and to initially explore the mechanism of CXCR4 on the invasion capability of esophageal carcinoma cell line EC9706 in vitro.【Methods】 RT-PCR were used to analyze the VEGF mRNA expression in EC9706 cells which transfected with two CXCR4 siRNA 48 h,EC9706 cells and EC9706 cells transfected with negative siRNA were used as control.Western blot and immunocytochemistry were used to analyze the VEGF protein expression in EC9706 cells which transfected with two CXCR4 siRNA 48 h,EC9706 cells and EC9706 cells transfected with negative siRNA were also used as control.【Results】 After transfected with two CXCR4 siRNA for 48 h,the results of RT-PCR showed that the expression of VEGF mRNA were decreased greatly than those in the negative control and blank control,and the differences were statistically significant（P 〈0.05）.The results of Western blot showed that CXCR4 siRNA could not only decrease the expression of VEGF mRNA but also could also decrease the expression in the protein level obviously,and the differences were statistically significant（P 〈0.05）.The results of Immunocytochemistry further manifested that CXCR4 siRNA could attenuate the protein expression of VEGF compared with those in the negative control and blank control,and the differences were statistically significant（P 〈0.05）.【Conclusion】 Specific blocking the expression CXCR4 can block the expression of VEGF too,so CXCR4 could probably take part in the invasion and metastasis of ESCC by controlling the expression of VEGF.CXCR4 is hopefully to be new target in cancer’s gene therapy.