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

目的:探讨NF-κBp65对食管鳞癌细胞EC9706上皮间质转化的影响。方法:NF-κBp65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-κBp65-ASODN转染前后EC9706细胞中NF-κBp65、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-κBp65-ASODN的EC9706细胞,其NF-κBp65 mRNA(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-κBp65-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-κBp65可能参与食管鳞状细胞癌的上皮转化。NF-κBp65-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。

Objective： To study the effect of NF-κBp65 on epithelial-mesenchymal transition （EMT） in esophageal squa- mous cell carcinoma cell line EC9706. Methods： Esophageal squamous cell carcinoma cell line EC9706 was transfected by chemically synthesized NF-KBp65-ASODN RT-PCR, immunocytochemistry and flow cytomertry were used to detect the mRNA and protein expression of NF-κBp65, E-cadherin and Vimentin in EC9706 before and after NF-KBp65-ASODN transfection. The morphological alterations of EC9706 cells were observed under inverted microscope and scarification test was used to detect the mobility of EC9706 cells before and after transfection. Results： After NF-κBp65-ASODN transfection, the mRNA （0.14〉0.06） and protein （immunocytochemistry： 11.33%＋1.40%, flow cytomertry： 11.78%） expressions of NF-κBp65 in EC9706 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-KBp65-ASODN transfection, the morphological alterations of EC9706 cells occurred and the migration length （0.45±0.08） was significantly shorter than that before transfection （0.81±0.11, P〈0.05）. Conclusion： NF-κBp65 may contribute to EMT in esophageal squamous cell carcinoma. NF-κBp65-ASODN transfection can in- hibit EMT in esophageal squamous cell carcinoma, up,regulate E-cadherin expression, down-regulate Vimentin expression and decrease cell mobility.