Relationship Between NF-κB and Epithelial-to-mesenchymal Transition of Radiation-induced Human Esophageal Carcinoma Cells
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摘要: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 mRNA表达明显下调（P=0.003），Vimentin、NF-κB mRNA表达升高（P=0.004，P=0.004）。相关性分析显示NF-κB与E-cadherin mRNA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。
Abstract:ObjectiveTo explore the relationship between NF-κB and epithelial-to-mesenchymal transition (EMT) of human esophageal carcinoma cells induced by the radiation.
MethodsWe 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-κB,E-cadherin and Vimentin in esophageal epithelial cell lines Eca109 and Eca109R60,respectively.
ResultsEca109R60 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-κB mRNA (P=0.004,P=0.004) . It showed a negative correlation between NF-κB 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.
ConclusionThe radio-resistance of human esophageal carcinoma cells Eca109R60 may be regulated through the interaction between EMT and NF-κB signaling pathway.
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Key words:
- Esophageal carcinoma /
- Radio-resistance /
- Radiotherapy /
- NF-κB /
- E-cadherin /
- Vimentin
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图 2 Eca109与Eca109R60细胞放射抵抗性的比较
Figure 2 Comparison of radio-resistance between Eca109 and Eca109R60 cells
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图 3 Eca109与Eca109R60细胞中E-cadherin mRNA、Vimentin mRNA及NF-κB mRNA的表达
Figure 3 Expression of E-cadherin mRNA,Vimentin mRNA and NF-κB mRNA in Eca109 and Eca109R60 cells
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图 4 Eca109与Eca109R60细胞中E-cadherin、Vimentin及NF-κBp65蛋白的表达 (×400)
Figure 4 Expression of E-cadherin,Vimentin and NF-κBp65 protein in Eca109 and Eca109R60 cells(×400)
表 1 Eca109与Eca109R60放射生物学参数
Table 1 Radiation biology parameter of Eca109 and Eca109R60 cells
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