特异性阻断Hedgehog信号通路对胰腺癌细胞凋亡的影响及分子机制

目的 观察特异性阻断Hedgehog信号通路对人胰腺癌Mia PaCa-2细胞凋亡的影响,探讨其可能的分子机制.方法 应用特异性Hedgehog通路阻断剂KAAD-eyelopamine阻断Mia PaCa-2细胞的Hedgehog信号通路,噻唑蓝(MTT)法检测阻断Hedgehog信号通路对胰腺癌细胞生长情况的影响;流式细胞技术观察细胞凋亡的变化;Western blot方法检测bax和bcl-2表达的变化.结果 KAAD-cyclopamine明显抑制胰腺癌Mia PaCa-2细胞生长,其作用呈剂量依赖性;阻断Hedgehog信号通路后,胰腺癌细胞凋亡显著增加,细胞中bax表达水平上调,bcl-2表达水平明显下调.结论 特异性阻断Hedgehog信号通路使胰腺癌细胞生长抑制,凋亡明显增加,其机制可能为通过上调bax及下调bcl-2的表达水平实现.

The effect and mechanism of blocking the hedgehog signaling pathway on apoptosis in pancreatic cancer Mia PaCa-2 cells

Objective To study the effect of blocking the hedgehog signaling pathway on apoptosis in pancreatic cancer Mid PaCa-2 cells and its possible mechanism.Methods The Mid PaCa-2 cells were treated with KAAD-cyclopamine at various concentrations for 48 h.Growth suppression was evaluated by MTT method.Flow cytometry (FCM) was used to assay the apoptotic rate.Western blot was used to detect the protein expression of bcl-2 and bax.Results A dose-dependent inhibitory effect of KAAD-cyclopamine on MiaPaCa-2 pancreatic caner cells was observed by MTT method.The result of FCM indicated that cancer cells treated with KAAD-cyclopamine for 48 h exhibited a significant increase of apoptosis,the expression of bcl-2 was significantly down-regulated and that of bax up-regulated.Conclusion KAAD-cyclopamine inhibits the proliferation of Mid PaCa-2 cells.Apoptostic induction may be the main antineoplastic mechanism,which is mediated by altering apoptosis-related genes through down-regulation of bcl-2 and up-regulation of bax proteins.