中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
8期
1683-1685
,共3页
张丽瑞%陈葳%赵乐%刘婷%李旭
張麗瑞%陳葳%趙樂%劉婷%李旭
장려서%진위%조악%류정%리욱
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
腫瘤壞死因子-α%覈轉錄因子-κB%上皮間質轉化%膀胱移行細胞癌%侵襲
종류배사인자-α%핵전록인자-κB%상피간질전화%방광이행세포암%침습
Tumor necrosis factor-α%Nuclear factor-kappa B%Epithelial-mesenchymal transition%Transitional cell carcinoma of bladder%Invasion
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 5 μg/L TNF-α处理BLX细胞,凝胶迁移实验(EMSA)观察NF-κB活性,Westem blot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.
目的 觀察腫瘤壞死因子-α(TNF-α)誘導人膀胱癌細胞的上皮間質轉化(EMT)及分子機製.方法 5 μg/L TNF-α處理BLX細胞,凝膠遷移實驗(EMSA)觀察NF-κB活性,Westem blot和免疫熒光檢測波形蛋白(Vimentin)和E-鈣黏蛋白(E-cadherin)的錶達及定位,體外侵襲轉移實驗觀察細胞侵襲轉移能力.逆轉錄-聚閤酶鏈反應(RT-PCR)檢測EMT相關轉錄因子的錶達.結果 TNF-α激活覈轉錄因子-κB(NF-κB),促使細胞由多邊形嚮紡錘形轉換,E-cadherin錶達下調(0.37±0.08比0.75±0.15,P＜0.05)且細胞膜定位消失,Vimentin錶達上調(0.46 ±0.12比0.00±0.00,P＜0.05),細胞轉移和侵襲能力提高(182.00±17.58比134.00 ±9.00；119.00±7.21比85.33±10.11,P＜0.05).TNF-α誘導Twist和Slug轉錄因子錶達上調(0.91 ±0.03比0.36±0.07；0.78±0.11比0.22±0.10,P＜0.01).結論 TNF-α激活NF-κB誘導膀胱癌細胞髮生EMT,可能在膀胱癌侵襲轉移中髮揮作用.
목적 관찰종류배사인자-α(TNF-α)유도인방광암세포적상피간질전화(EMT)급분자궤제.방법 5 μg/L TNF-α처리BLX세포,응효천이실험(EMSA)관찰NF-κB활성,Westem blot화면역형광검측파형단백(Vimentin)화E-개점단백(E-cadherin)적표체급정위,체외침습전이실험관찰세포침습전이능력.역전록-취합매련반응(RT-PCR)검측EMT상관전록인자적표체.결과 TNF-α격활핵전록인자-κB(NF-κB),촉사세포유다변형향방추형전환,E-cadherin표체하조(0.37±0.08비0.75±0.15,P＜0.05)차세포막정위소실,Vimentin표체상조(0.46 ±0.12비0.00±0.00,P＜0.05),세포전이화침습능력제고(182.00±17.58비134.00 ±9.00；119.00±7.21비85.33±10.11,P＜0.05).TNF-α유도Twist화Slug전록인자표체상조(0.91 ±0.03비0.36±0.07；0.78±0.11비0.22±0.10,P＜0.01).결론 TNF-α격활NF-κB유도방광암세포발생EMT,가능재방광암침습전이중발휘작용.
Objective To investigate the effects of nuclear factor-kappaB (NF-κB) activity induced by tumor necrosis factor-αt (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 examined 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 immunofluorescent assay.In vitro migration and invasion abilities were determined by using Millicell assay.EMT-related molecules were examined by using reverse transcription-polymerase chain reaction (RT-PCR).Results TNF-α could induce NF-κB activity in BLX cells,promoting the converstion of BLX cells from rounded epithelial-like cells to fibroblast-like cells with elongated morphologies.The BLX cells treated with TNF-α showed 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).TNFoα-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,theupregulation of Twist and Slug was demonstrated in TNFα-treated BLX cells 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 suppressing the metastasis and invasion of the bladder carcinoma.
