盐酸去氢骆驼蓬碱诱导人胰腺癌AsPC-1细胞凋亡

摘要

Aim:To investigate the inhibitory effect of Harmine hydrochloride on human pancreatic cancer cell line AsPC-1 and the possible mechanism.Methods:MTT and cell colony formation inhibitory assay were used to investigate the inhibition effect of Harmine hydrochloride on human pancreatic cancer cell line AsPC-1.Changes in apoptotic cell percentage were calculated by flow cytometry.Western blotting assay was used to determine cell apoptosis and autophagy.Results:MTT showed that Harmine hydrochloride could inhibited human pancreatic cancer AsPC-lcells in a dose-dependent manner.IC50 of Harmine hydrochloride on AsPC-1 cells at 24 h,48 h and 72 h was 116.5 μmol · L-1,66 μmol · L-1,22.3 μmol · L-1 respectively;Cell clone formation inhibition assay demonstrated that cell clones were decreased with increase of the concentration of Harmine hydrochloride.Annexin V-FITC/PI staining also showed a dose-dependent effect on late apoptotic and necrotic cells.Western blotting assay showed that the apoptosis and autophagy landmark protein Caspase-3,PARP,LC3-II and P62 protein expression levels changed gradually with increasing of Harmine hydrochloride.Conclusion:Harmine hydrochloride inhibits the cell survival of human pancreatic cancer cell AsPC-1 in a dose-dependent manner.Harmine hydrochloride inhibits cell growth by inducing apoptotic and autophagy.%目的:探讨盐酸去氢骆驼蓬碱对人胰腺癌AsPC-1细胞的增殖抑制作用及其可能的作用机制.方法:采用噻唑蓝(MTF)法及克隆形成抑制实验观察盐酸去氢骆驼蓬碱对胰腺癌AsPC-1细胞的增殖抑制作用,流式细胞仪检测细胞凋亡水平,Western blotting检测细胞凋亡及自噬相关蛋白的表达水平.结果:盐酸去氢骆驼蓬碱作用人胰腺癌AsPC-1细胞后,不同浓度的药物对其生长均有抑制作用,且呈剂量-效应关系(P＜0.01),24、48及72h的IC50分别约为116.5、66、22.3 μmol·L-1；与对照组相比随着盐酸去氢骆驼蓬碱浓度的增加,细胞克隆逐渐减少；不同浓度盐酸去氢骆驼蓬碱作用后,均出现明显的晚期凋亡及坏死细胞群,且呈剂量-效应关系；盐酸去氢骆驼蓬碱作用后,胰腺癌AsPC-1细胞出现Caspase-3、PARP酶切活化,同时自噬标记性蛋白LC3Ⅱ增加,P62减少.结论:盐酸去氢骆驼蓬碱通过诱导细胞凋亡及自噬的方式抑制人胰腺癌AsPC-1细胞的生长.