转化生长因子β1与食管鳞状细胞癌上皮间质转化的关系  

Transforming growth factor β1 regulation of epithelial-mesenchymal transition in esophagus squamous cell carcinoma

作　　者：孙洋[1] 李珊珊[1] 王新华[1] 王小军[1] 阎爱华[2] 

机构地区：[1]郑州大学第一附属医院病理科河南省肿瘤病理重点实验室,450052 [2]郑州大学基础医学院癌前研究室

出　　处：《中华病理学杂志》2008年第8期542-548,共7页Chinese Journal of Pathology

基　　金：河南省重点科技攻关项目（072102310042）

摘　　要：目的探讨食管鳞状细胞癌中转化生长因子β1（TGFβ1）与上皮间质转化的关系以及阻断TGFβ1通路对食管鳞状细胞癌上皮间质转化的影响。方法将化学合成的TGFβ1反义寡核苷酸（TGFβ1—ASODN）转染食管鳞状细胞癌细胞株EC9706，采用RT—PCR、免疫细胞化学及流式细胞术检测阻断前后对TGFβ1活性的影响以及对上皮性标志物E—cadherin及间质性标志物波形蛋白表达的影响；观察TGFβ1—ASODN转染前后细胞形态学的变化；采用细胞划痕试验检测TGFβ1—ASODN转染前后细胞迁移能力的变化。结果转染TGFβ1—ASODN可有效的抑制EC9706细胞中TGFβ1的活性，其mRNA（0．25±0．07）及蛋白（35．07％±1．42％）的表达均明显低于转染前mRNA（0．43±0．09）及蛋白（43．57％±1．77％）的水平（Х^2=13．847及x。=84．120，均P〈0．05）；并提高E—cadherin的表达，抑制波形蛋白的表达。转染后E—cadherin mRNA（0．38±0．09）及蛋白（17．13％±1．45％）的表达明显高于转染前mRNA（0．22±0．06）及蛋白（12．53％±1．31％）的水平（Х^2=0．160及Х^2=40．008，均P〈0．05）；波形蛋白mRNA（0．73±0．07）及蛋白（14．15％±1．46％）的表达低于转染前mRNA（0．89±0．09）及蛋白（17．97％±1．42％）水平（Х^2=0．160及Х^2=21．103，均P〈0．05）。TGFβ1-ASODN转染可明显抑制细胞的运动能力，转染后细胞迁移距离（0．45±0．05）比转染前（0．81±0．11）明显缩小（Х^2=16．854，P〈0．05）。结论TGFβ1可能参与了食管鳞状细胞癌的上皮间质转化，TGFβ1—ASODN可导致食管鳞状细胞癌细胞上皮性标志物E—cadherin表达上调、间质性标志物波形蛋白表达下调、形态发生改变以及细胞移动能力降低，阻断TGFN信号可抑制食管鳞状细胞癌细胞的上皮间质转化。Objective To study the functional role of transforming growth factor β1 （ TGFβ1 ） in the regulation of epithelial-mesenchymal transition （EMT） and the effect of TGFβ1-ASODN blockage of EMT in esophagus squamous cell carcinoma. Methods Esophageal squamous cell carcinoma cell line EC9706 was transfected with chemically synthesized TGFβ1-ASODN. RT-PCR, immunohistochemistry and flow cytometry were used to detect the protein and mRNA expressions of TGF-β1, E-cadherin and vimentin before and after the transfection. Morphological changes were documented and scarification test was used to detect the migration potential of EC9706 before and after the transfection. Results After TGFβ1-ASODN transfection, mRNA （0. 25 ± 0. 07 ） and protein （35.07% ± 1.42% ） expressions of TGFβ1 in EC9706 were significantly lower than those before transfection （mRNA： 0. 43 ±0. 09; protein： 43. 57% ± 1.77%, Х^2 = 13. 847 and Х^2=84.120, P〈0.05）. The mRNA （0.38±0.09） and protein （17.13% ±1.45%） expressions of E-cadhefin were significantly higher than those before transfection （0. 22 ±0.06; 12. 53% ± 1.31%,Х^2 = 0.160 and Х^2 =40.008,P〈0.05） and the mRNA （0.73 ±0.07） and protein （14.15% ± 1.46%） expressions of vimentin were significantly lower than those （ 0. 89 ±0.09 ; 17.97% ± 1.42% ） before transfection （Х^2 =0. 160 and Х^2 =21. 103,P 〈0. 05）. Scarification test showed that after transfection, the mobility of EC9706 was significantly inhibited and its migration length （0. 45 ±0. 05 ） was significantly shorter than that before the transfection（0. 81 ± 0. 11, Х^2 = 16. 854, P 〈 0. 05）. Conclusions TGFβ1 may contribute to EMT in esophageal squamous cell carcinoma. TGFβ1-ASODN leads to an over-expression of E-cadherin and a down-regulation of vimentin, along with the morphological alterations and migration inhibition, indicating that a blockage of TGFβ1 suppresses EMT in esophagus squamous cell carcinoma.

关 键 词：食管肿瘤 转化生长因子Β1 转化 遗传 

分 类 号：R686[医药卫生—骨科学]