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

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-cadherin、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 T24,5637 and J82 cells which treated for 48 hours using γ-secretase inhibitor by inverted microscope. The mRNA and protein levels 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 proliferation 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 invasion 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.