摘要
目的观察肿瘤坏死因子-α（TNF-α）诱导人膀胱癌细胞的上皮间质转化（EMT）及分子机制。方法5μg／LTNF-α处理BLX细胞，凝胶迁移实验（EMSA）观察NF-κB活性，Westernblot和免疫荧光检测波形蛋白（Vimentin）和E-钙黏蛋白（E-cadherin）的表达及定位，体外侵袭转移实验观察细胞侵袭转移能力。逆转录-聚合酶链反应（RT-PCR）检测EMT相关转录因子的表达。结果TNF-α激活核转录因子-κB（NF-κB），促使细胞由多边形向纺锤形转换，E-cadherin表达下调（0．37±0．08比0．75±0．15，P〈0．05）且细胞膜定位消失，Vimentin表达上调（0．46±0．12比0．00±0．00，P〈0．05），细胞转移和侵袭能力提高（182．00±17．58比134．00±9．00；119．00±7．21比85．33±10．11，P〈0．05）。TNF-α诱导Twist和Slug转录因子表达上调（0．91±0．03比0．36±0．07；0．78±0．11比0．22±0．10，P〈0．01）。结论TNF-α激活NF-κB诱导膀胱癌细胞发生EMT，可能在膀胱癌侵袭转移中发挥作用。
Objective To investigate the effects of nuclear factor-kappaB （NF-κB） activity in- duced by tumor necrosis factor-α （TNF-α） on epithelial-mesenchymal transition （EMT） of bladder cancer cells. Methods BLX cells were treated with TNF-α （5 μg/L） and the morphological changes were exam- ined by phase contrast microscope. NF-κB activiy of BLX cells was measured by electrophoretic mobility shift assay （EMSA）. Vimentin and E-cadherin were detected by using Western blotting and immunofluo- rescent assay. In vitro migration and invasion abilities were determined by using Millice11 assay. EMT-relat- ed molecules were examined by using reverse transcription-polytnerase chain reaction （RT-PCR）. Results TNF-α could induce NF-κB activity in BLX cells, promoting the converstion of BLX cells from rounded ep- ithelial-like cells to fibroblast-like cells with elongated morphologies. The BLX cells treated with TNF-αshowed the downregulation of E-cadherin protein （ 0. 37 ± 0. 08 vs. 0. 75 ± 0. 15, P 〈 0. 05 ）, especially membrane localization loss, and upregulation of vimentin protein （ 0. 46 ± 0. 12 vs. 0. 00 ± 0. 00, P 〈 0. 05）. TNFα-treated BLX cells showed more motile and invasive than untreated control cells （182. 00 ± 17.58 vs. 134. 00 ± 9. 00 ; 119.00 ± 7.21 vs. 85.33 ± 10. 11, P 〈 0. 05）. Moreover, the upregulation of Twist and Slug was demonstrated in TNFα-treated BLX ceils compared to control cells （0. 91 ± 0. 03 vs. 0. 36 ± 0. 07 ; 0. 78 ± 0. 11 vs. 0. 22 ± 0. 10, P 〈 0.01 ）. Conclusion Activation of NF-κB induced by TNF-α promotes EMT in bladder cancer cells, suggesting that NF-κB may also be a potential target for sup- pressing the metastasis and invasion of the bladder carcinoma.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第8期1683-1685,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30973429、81202055）
关键词
肿瘤坏死因子-Α
核转录因子-ΚB
上皮间质转化
膀胱移行细胞癌
侵袭
Tumor necrosis factor-α
Nuclear factor-kappa B
Epithelial-mesenchymal transi- tion
Transitional cell carcinoma of bladder
Invasion
