Aurora-A inhibitor 1对人胰腺癌细胞增殖、凋亡的影响

Effect of Aurora-A inhibitor 1 on proliferation;apoptosis of PANC-1 cells

目的检测Aurora-A在胰腺癌中的表达，并探讨特异性抑制剂Aurora-A inhibitor 1在胰腺癌增殖、凋亡中的作用。方法利用Western blot检测人类正常胰腺上皮细胞（HPNE）和胰腺癌细胞（PANC-1）中Aurora-A、p-Aurora-A蛋白的表达。细胞计数试剂盒（CCK-8）技术检测Aurora-A inhibitor 1对胰腺癌细胞增殖的影响；流式细胞术检测细胞周期以及凋亡，Western blot方法检测p-Aurora-A、Aurora-A、细胞周期和凋亡相关蛋白的表达。结果Western blot结果示较HPNE细胞，PANC-1细胞中Aurora-A蛋白表达量上调22.067倍，p-Aurora-A蛋白表达量上调8.962倍。CCK-8结果示：Aurora-A inhibitor 1可明显抑制PANC-1细胞的增殖（P=0.019）。流式细胞周期检测显示：PANC-1细胞经过Aurora-A inhibitor 1处理后，G1/S期细胞明显减少，G2/M细胞明显增加[空白组（9.6±2.3）%、对照组（7.5±4.8）%、实验组（61.2±4.0）%]；同时，p21蛋白表达水平明显升高（空白组：1.000±0.000、对照组：1.671±0.111、实验组：3.208±0.284）。凋亡分析：实验组较其他两组PANC-1细胞凋亡明显增加（空白组：1.000±0.000、对照组1.053±0.108、实验组1.902±0.276），裂解的半胱氨酰天冬氨酸特异性蛋白酶（Cleaved Caspase-3）蛋白表达水平明显增加（空白组：1.000±0.000、对照组：1.274±0.112、实验组：1.754±0.062）。结论Aurora-A、p-Aurora-A在胰腺癌中明显高表达，特异性抑制剂Aurora-A inhibitor 1通过抑制p-Aurora-A将胰腺癌细胞明显阻滞于G2/M期，并可抑制胰腺癌增殖，诱导细胞凋亡。

ObjectiveTo investigate the expression of Aurora kinase A in normal human pancreatic epithelial cell and pancreatic cancer cell lines, and explore the role of Aurora-A inhibitor 1 on proliferation, apoptosis of pancreatic cancer cell PANC-1.MethodsThe expression level of Aurora-A and p-Aurora-A protein in two different cells was detected using Western blotting. Cell counting kit-8 （CCK-8） was constructed to investigate cell proliferation. Flow cytometry （FCM） and Western blotting were used to detect cell cycle and cell apoptosis.ResultsWestern blotting indicated that Aurora-A and p-Aurora-A were over-expression in pancreatic cancer. The results of CCK-8 showed that Aurora-A could inhibit the proliferation of PANC-1 （P=0.019）. The test of FCM showed that Aurora-A inhibitor arrest the cells during G2/M [Blank： （9.6±2.3）%, dimethyl sulfoxide （DMSO）： （7.5±4.8）%, Aurora-A inhibitor 1： （61.2±4.0）%] and Western blotting further showed p21 was up-regulated （Blank： 1.000±0.000, DMSO： 1.671±0.111, Aurora-A inhibitor 1： 3.208±0.284）. Moreover, FCM indicated that the apoptosis was significantly increased in Aurora-A inhibitor 1 group after treatment （Blank： 1.000±0.000, DMSO： 1.053±0.108, Aurora-A inhibitor 1： 1.902±0.276）. The results of Western blotting showed the expression of Cleavage of cysteinyl aspartate specific protease （Cleaved Caspase-3） was raised （Blank： 1.000±0.000, DMSO： 1.274±0.112, Aurora-A inhibitor 1： 1.754±0.062）.ConclusionAurora-A and p-Aurora-A is over-expression in pancreatic cancer. Inhibition of p-Aurora-A by Aurora-A inhibitor 1 can arrest pancreatic cancer cell in G2/M. Moreover, Aurora-A inhibitor 1 decrease proliferation and promote cell apoptosis.