Notch信号通路调控上皮-间质转化影响膀胱癌侵袭性和耐药性OACSCDCSTPCD
Notch signaling pathway regulates epithelial-mesenchymal transition and affects the invasiveness and drug resistance of bladder cancer
目的:体外探讨Notch信号通路对膀胱癌细胞侵袭性与耐药性的影响及分子机制。方法采用Notch信号通路受体完全阻断剂(γ分泌酶抑制剂)处理膀胱癌T24、5637和J82细胞48 h后,倒置显微镜观察膀胱癌细胞增生及形态;用RT-PCR和Western blot在mRNA和蛋白水平检测上皮-间质转化( EMT)分子标志物E-cadherin、N-cad-herin、vimentin和Alpha-smooth muscle actin的表达;MTT、Transwell检测膀胱癌细胞耐药性及侵袭能力。结果完全阻断Notch信号通路后,镜下显示膀胱癌细胞形态变小,细胞分散;EMT分子标志物E-cadherin mRNA和蛋白水平表达上调( P<0.05),N-cadherin、vimentin、Alpha-smooth muscle actin mRNA 和蛋白水平表达下调( P<0.05);膀胱癌细胞T24、5637和J82增殖明显被抑制( P<0.05);膀胱癌细胞T24、5637和J82穿过微孔膜的细胞数明显减少( P<0.05)。结论 Notch信号通路可通过调控EMT的变化而改变膀胱癌的侵袭性与耐药性。
Objective To investigate the effect of notch signaling pathway on drug resistance and invasiveness of bladder cancer .Methods We observed the changes of growth and morphology of bladder cancer T 24 , 5637 and J82 cells which treated for 48 hours using γ-secretase inhibitor by inverted microscope .The mRNA and protein lev-els of the EMT molecular markers , including E-cadherin , N-cadherin , vimentin and Alpha-smooth muscle actin were examined by RT-PCR and Western blot in bladder cancer cells;Detected the changes of drug resistance and invasion respectively by MTT and Transwell in bladder cancer cells .Results After completely blocking the Notch signaling pathway , the inverted microscope showed that bladder cancer cells became smaller and more disperse ;RT-PCR and Western blot showed the mRNA and protein levels of E-cadherin were up-regulated ( P<0.05 ) , contrast , N-cadherin , vimentin and Alpha-smooth muscle actin were down-regulated ( P<0.05 ); The prolifera-tion of bladder cancer cells were significantly inhibited by MTT test;The number of through microporous membrane cells significantly decreased ( P<0.05 ) shown by Transwell test .Conclusions The Notch signaling pathway is completely blocked that nhibites proliferation and EMT of bladder cancer cells , reduces drug resistance and inva-sion in bladder cancer cells .It suggests that drug resistance and invasiveness of bladder cancer can be changed through EMT which is regulated through notch signaling pathway .
刘志欢;王义兵;王共先;黄亮;郎斌;许晓源;傅斌
南昌大学第一附属医院泌尿外科,江西南昌330006南昌大学第一附属医院泌尿外科,江西南昌330006南昌大学第一附属医院泌尿外科,江西南昌330006 江西省泌尿外科研究所,江西南昌330006南昌大学第一附属医院泌尿外科,江西南昌330006澳门理工学院高等卫生学校,澳门999078九江学院基础医学院江西省生物医药重点实验室,江西九江332000
医药卫生
膀胱癌Notch信号通路上皮-间质转化γ分泌酶抑制剂
bladder cancerNotch signalepithelial-mesenchymal transitionγ-secretase inhibitor
《基础医学与临床》 2015 (2)
145-151,7
国家自然科学基金(811606272);江西省自然科学基金(800GZY0039);江西省青年科学家基金(2010JX02761);澳门科学技术发展基金(064/2012/A)
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