NF-κB与放射线诱导的人食管癌细胞上皮间质转化间的关系

Relationship Between NF-κB and Epithelial-to-mesenchymal Transition of Radiationinduced Human Esophageal Carcinoma Cells

目的探讨NF-κB与放射线诱导的人食管癌细胞上皮间质转化(EMT)间的关系。方法累积照射食管癌Eca109细胞60 Gy,得到Eca109R60细胞,克隆形成实验检测其放射抵抗性,显微镜下观察细胞形态学变化。Real-time PCR和免疫细胞化学检测Eca109和Eca109R60细胞NF-κB、E-cadherin和Vimentin的表达情况。结果 Eca109R60细胞具备了EMT样表型且放射抵抗性更强。Eca109R60细胞中E-cadherin m RNA表达明显下调(P=0.003),Vimentin、NF-κB m RNA表达升高(P=0.004,P=0.004)。相关性分析显示NF-κB与E-cadherin m RNA表达呈负相关,与Vimentin未见有明显相关性,但已显示出正相关的趋势。免疫细胞化学法检测Eca109R60细胞E-cadherin阳性染色程度明显减弱,Vimentin、NF-κB p65阳性染色程度明显增强。结论 Eca109R60细胞放射抗性可能由EMT与NF-κB信号通路相互作用调控。

Objective To explore the relationship between NF-r,B and epithelial-to-mesenchymal transition （EMT） of human esophageal carcinoma cells induced by the radiation. Methods We irradiated cumulatively human esophageal carcinoma cells Eca109 with 60Gy to get Eca109R60 cells. The radio-resistance was detected by means of a standard colony-formation assay. Cells morphological change was observed under microscope. Meanwhile, Real-time PCR and immunocytochemistry were used to detect the expression level of NF-r,B, E-cadherin and Vimentin in esophageal epithelial cell lines Eca109 and Eca109R60, respectively. Results Eca109R60 cells had EMT-like phenotype and stronger irradiation resistance（IR）. In Eca109R60 cells, the expression level of E-cadherin mRNA was significantly down-regulated （P=0.003）. There were significant upregulation in the expression levels of Vimentin and NF-tcB mRNA （P=0.004, P=0.004）. It showed a negative correlation between NF-r,B and E-cadherin by the correlation analysis. There was not an evident correlation between NF-κB and Vimentin, but a trend to negative correlation. From the result of immunocytochemistry staining, a weaker expression of E-cadherin protein while stronger expression of Vimentin and NF-κB p65 protein were found. Conclusion The radio-resistance of human esophageal carcinoma cells Eca109R60 may be regulated through the interaction between EMT and NF-κB signaling pathway.