中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2012年
9期
749-752
,共4页
皮雷鸣%刘勇%余长云%蔡耿明%黄东海%邱元正%田勇泉%张欣
皮雷鳴%劉勇%餘長雲%蔡耿明%黃東海%邱元正%田勇泉%張訢
피뢰명%류용%여장운%채경명%황동해%구원정%전용천%장흔
头颈部肿瘤%癌,鳞状细胞%儿茶素%转化生长因子β1%上皮间质转化
頭頸部腫瘤%癌,鱗狀細胞%兒茶素%轉化生長因子β1%上皮間質轉化
두경부종류%암,린상세포%인다소%전화생장인자β1%상피간질전화
Head and neck neoplasms%Carcinoma,squamous cell%Catechin%Transforming growth factor beta 1%Epithelial-mesenchymal transition
目的 探讨表没食子儿茶素没食子酸酯(epigallocaechin-3-gallate,EGCG)对重组人转化生成因子β1( transforming growth factor-β1,TGF-β1)蛋白诱导的头颈鳞癌细胞上皮-间质转化的影响及其分子机制.方法 5 ng/ml TGF-β1作用于头颈鳞癌Tu686细胞24 h后,再加入浓度为20 μmol/L的EGCG处理24 h,观察其上皮-间质转化的形态学改变;5 ng/ml TGF-β1作用24 h后,加入不同浓度(0、10、20、30 μmol/L) EGCG作用24 h,反转录聚合酶链反应(RT-PCR)和Western blot检测上皮细胞标志物钙黏蛋白(E-cadherin)、间质细胞标志物波形蛋白(Vimentin)及TGF-β1信号通路抑制性蛋白Smad7的表达;20 μmol/L EGCG作用于5 ng/ml TGF-β1处理过的Tu686细胞,于不同的时间点(6、12、24 h)收集细胞,检测E-cadherin及Vimentin的表达.结果 TGF-β1能诱导头颈鳞癌Tu686细胞发生上皮-间质转化形态学及分子水平改变,E-cadherin表达下调、Vimentin的表达增强,TGF-β1信号通路抑制性蛋白Smad7表达下调;EGCG能有效抑制TGF-β1诱导的上皮-间质转化过程,能促进E-cadherin表达上调、Vimentin表达下降及Smad7表达升高.结论 EGCG能有效抑制TGF-β1诱导的头颈鳞癌Tu686细胞发生的上皮-间质转化过程,且该抑制作用可能与TGF-β1信号通路中抑制性因子Smad7的表达升高有关.
目的 探討錶沒食子兒茶素沒食子痠酯(epigallocaechin-3-gallate,EGCG)對重組人轉化生成因子β1( transforming growth factor-β1,TGF-β1)蛋白誘導的頭頸鱗癌細胞上皮-間質轉化的影響及其分子機製.方法 5 ng/ml TGF-β1作用于頭頸鱗癌Tu686細胞24 h後,再加入濃度為20 μmol/L的EGCG處理24 h,觀察其上皮-間質轉化的形態學改變;5 ng/ml TGF-β1作用24 h後,加入不同濃度(0、10、20、30 μmol/L) EGCG作用24 h,反轉錄聚閤酶鏈反應(RT-PCR)和Western blot檢測上皮細胞標誌物鈣黏蛋白(E-cadherin)、間質細胞標誌物波形蛋白(Vimentin)及TGF-β1信號通路抑製性蛋白Smad7的錶達;20 μmol/L EGCG作用于5 ng/ml TGF-β1處理過的Tu686細胞,于不同的時間點(6、12、24 h)收集細胞,檢測E-cadherin及Vimentin的錶達.結果 TGF-β1能誘導頭頸鱗癌Tu686細胞髮生上皮-間質轉化形態學及分子水平改變,E-cadherin錶達下調、Vimentin的錶達增彊,TGF-β1信號通路抑製性蛋白Smad7錶達下調;EGCG能有效抑製TGF-β1誘導的上皮-間質轉化過程,能促進E-cadherin錶達上調、Vimentin錶達下降及Smad7錶達升高.結論 EGCG能有效抑製TGF-β1誘導的頭頸鱗癌Tu686細胞髮生的上皮-間質轉化過程,且該抑製作用可能與TGF-β1信號通路中抑製性因子Smad7的錶達升高有關.
목적 탐토표몰식자인다소몰식자산지(epigallocaechin-3-gallate,EGCG)대중조인전화생성인자β1( transforming growth factor-β1,TGF-β1)단백유도적두경린암세포상피-간질전화적영향급기분자궤제.방법 5 ng/ml TGF-β1작용우두경린암Tu686세포24 h후,재가입농도위20 μmol/L적EGCG처리24 h,관찰기상피-간질전화적형태학개변;5 ng/ml TGF-β1작용24 h후,가입불동농도(0、10、20、30 μmol/L) EGCG작용24 h,반전록취합매련반응(RT-PCR)화Western blot검측상피세포표지물개점단백(E-cadherin)、간질세포표지물파형단백(Vimentin)급TGF-β1신호통로억제성단백Smad7적표체;20 μmol/L EGCG작용우5 ng/ml TGF-β1처리과적Tu686세포,우불동적시간점(6、12、24 h)수집세포,검측E-cadherin급Vimentin적표체.결과 TGF-β1능유도두경린암Tu686세포발생상피-간질전화형태학급분자수평개변,E-cadherin표체하조、Vimentin적표체증강,TGF-β1신호통로억제성단백Smad7표체하조;EGCG능유효억제TGF-β1유도적상피-간질전화과정,능촉진E-cadherin표체상조、Vimentin표체하강급Smad7표체승고.결론 EGCG능유효억제TGF-β1유도적두경린암Tu686세포발생적상피-간질전화과정,차해억제작용가능여TGF-β1신호통로중억제성인자Smad7적표체승고유관.
Objective To study the effect and molecular mechanism of epigallocaechin-3-gallate (EGCG) on epithelial-mesenchymal transition (EMT) in vitro induced by human recombinant TGF-β1 protein in squamous cell carcinoma of the head and neck.Methods EMT morphological changes of Tu686 cells were observed after sequential treatment of 5 ng/ml TGF-β1 and 20 μmol/L EGCG.Tu686 cells were collected after the treatment of 5 ng/ml TGF-β1 for 24 h and EGCG with different concentrations (0,10,20,30 μmol/L) for another 24 h or 20 μmol/L EGCG treatment for different time phase(6,12,24 h).Then RT-PCR and Western-blot were applied to detect mRNA and protien expression level of epithelial cell marker E-cadherin,mesenchymal cell marker Vimentin and Smad7,an inhibit molecule of TGF-β1 mediated pathway in Tu686 cells.Results TGF-β1 successfully induced characterized EMT morphological and molecular changes in Tu686 cells,in which expression of E-cadherin decreased,Vimentin increased and Smad7 declined.However,EGCG could reverse the TGF-β1 mediated process of EMT by downregulating the expression of Vimentin and upregulating the expression of E-cadherin and Smad7.Conclusion EGCG significantly inhibits TGF-β1-mediated EMT inTu686 cell lines of SCCHN,which maybe associated with the upregulated-expression of Smad7,an inhibitor in TGF-β1 signaling pathway.
