张红燕, 王小军, 王 丰, 王新华, 孙 洋, 李珊珊. NF-κBp65对食管鳞癌细胞上皮间质转化的影响*[J]. 中国肿瘤临床, 2010, 37(4): 187-189. DOI: 10.3969/j.issn.1000-8179.2010.04.003
引用本文: 张红燕, 王小军, 王 丰, 王新华, 孙 洋, 李珊珊. NF-κBp65对食管鳞癌细胞上皮间质转化的影响*[J]. 中国肿瘤临床, 2010, 37(4): 187-189. DOI: 10.3969/j.issn.1000-8179.2010.04.003
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ZHANG Hongyan, WANG Xiaojun, WANG Feng, WANG Xinhua, SUN Yang, LI Shanshan. Effect of NF-κBp65 on Epithelial-mesenchymal Transition in Esophageal Squmous Cell Carcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(4): 187-189. DOI: 10.3969/j.issn.1000-8179.2010.04.003
Citation: ZHANG Hongyan, WANG Xiaojun, WANG Feng, WANG Xinhua, SUN Yang, LI Shanshan. Effect of NF-κBp65 on Epithelial-mesenchymal Transition in Esophageal Squmous Cell Carcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(4): 187-189. DOI: 10.3969/j.issn.1000-8179.2010.04.003
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NF-κBp65对食管鳞癌细胞上皮间质转化的影响*

Effect of NF-κBp65 on Epithelial-mesenchymal Transition in Esophageal Squmous Cell Carcinoma

    摘要
  • 摘要: 目的：探讨NF-κ Bp65对食管鳞癌细胞EC9706上皮间质转化的影响。方法：NF-κ Bp65反义寡核苷酸（NF-κBp65-ASODN ）转染食管鳞癌细胞EC9706，采用RT-PCR、免疫细胞化学及流式细胞术检测NF-κ Bp65-ASODN 转染前后EC9706细胞中NF-κ Bp65、上皮性标志物E-cadherin 及间质性标志物Vimentin mRNA 及蛋白表达的变化，观察转染前后细胞形态学的改变，细胞划痕实验检测转染前后细胞运动能力的变化。结果：转染NF-κ Bp65-ASODN 的EC9706细胞，其NF-κ Bp65mRNA（0.14±0.06）及蛋白（免疫细胞化学：11.33± 1.40% ，流式细胞术：11.78%）的表达低于转染前（mRNA ：0.45± 0.05；蛋白：免疫细胞化学：17.17± 1.66% ，流式细胞术：15.76%）（P<0.05），上皮性标志物E-cadherin mRNA（0.36± 0.08）及蛋白（免疫细胞化学：17.58± 1.86% ，流式细胞术：14.98%）的表达高于转染前（mRNA ：0.22± 0.06；蛋白：免疫细胞化学：14.42± 1.63% ，流式细胞术：12.22%）（P<0.05），间质性标志物Vimentin mRNA（0.75± 0.07）及蛋白（免疫细胞化学：16.00± 1.41% ，流式细胞术：12.90%）的表达低于转染前（mRNA ：0.89± 0.09；蛋白：免疫细胞化学：19.50± 0.89% ，流式细胞术：17.76%）（P<0.05）。 转染NF-κ Bp65-ASODN 后，EC9706细胞形态发生改变，细胞迁移距离（0.45± 0.08）较转染前（0.81± 0.11）明显缩短（P<0.05）。 结论：NF-κ Bp65可能参与食管鳞状细胞癌的上皮转化。NF-κ Bp65-ASODN 转染可抑制食管鳞癌细胞上皮间质转化，上皮性标志物E-Cadherin 表达上调、间质性标志物Vimentin 表达下调，且细胞形态发生改变、细胞运动能力降低。

     

    Abstract: Objective:To study the effect of NF-κ Bp65on epithelial-mesenchymal transition (EMT) in esophageal squa-mous cell carcinoma cell line EC9706. Methods:Esophageal squamous cell carcinoma cell line EC9706was transfected by chemical ly synthesized NF-κ Bp65-ASODN. RT-PCR, immunocytochemistry and flow cytomertry were used to detect the mRNA and protein expression of NF-κ Bp65, E-cadherin and Vimentin in EC 9706before and after NF-κ Bp65-ASODN trans -fection. The morphological alterations of EC 9706cells were observed under inverted microscope and scarification test was used to detect the mobility of EC 9706cells before and after transfection. Results:After NF-κ Bp65-ASODN transfection, the mRNA (0.14± 0.06) and protein (immunocytochemistry: 11.33%±1.40%, flow cytomertry:11.78% ) expressions of NF-κ Bp65 in EC 9706 were significantly lower than those before transfection (mRNA: 0.45± 0.05; protein: immunocytochemistry: 17.17% ±1.66% , flow cytomertry: 15.76% ,P<0.05), the mRNA ( 0.36± 0.08) and protein (immunocytochemistry: 17.58% ± 1.86% , flow cytomertry: 14.98% ) expressions of E-cadherin in EC9706 cells were significantly higher than those before transfection (mRNA: 0.22± 0.06; protein: immunocytochemistry: 14.42%±1.63%, flow cytomertry: 12.22%, P<0.05) and the mRNA (0.75± 0.07) and protein (immunocytochemistry: 16.00%±1.41%, flow cytomertry: 12.90%) expressions of Vimentin were significantly lower than those before transfection (mRNA: 0.89± 0.09; protein: immunocytochemistry: 19.50± 0.89% , flow cytomertry: 17.76%,P<0.05). After NF-κ Bp65-ASODN transfection, the morphological alterations of EC 9706 cells oc-curred and the migration length ( 0.45± 0.08) was significantly shorter than that before transfection ( 0.81± 0.11, P<0.05). Conclusion: NF-κ Bp65may contribute to EMT in esophageal squamous cel l carcinoma. NF-κ Bp65-ASODN transfection can in -hibit EMT in esophageal squamous cell carcinoma, up-regulate E-cadherin expression, down-regulate Vimentin expression and decrease cell mobility.

     

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