阿司匹林对胰腺癌细胞上皮间质转化和吉西他滨耐药性的影响

Effects of aspirin on epithelial mesenchymal transition and gemcitabine resistance of human pancreatic cancer cells

目的探讨阿司匹林对人胰腺癌细胞上皮间质转化(EMT)和吉西他滨耐药性的影响。方法应用MTT法检测阿司匹林(0、1、2、5、10mmol/L)对三种胰腺癌细胞(SW1990、PATU-8988、PANC-1)生存率的影响以及阿司匹林(2mmol/L)与吉西他滨(0.05mg/L)联合用药对SW1990细胞增殖的影响。平板克隆实验检测药物对克隆形成能力的影响;流式细胞术检测细胞凋亡情况;细胞划痕实验观察细胞的迁移能力;Western blot检测凋亡相关蛋白及EMT相关蛋白表达水平。结果(1)与对照组相比,低浓度阿司匹林(1、2mmol/L)对PATU-8988、PANC-1有生长抑制作用(P<0.05、P<0.01);高浓度阿司匹林(5、10mmol/L)对SW1990、PATU-8988、PANC-1均有较明显的生长抑制作用(P<0.05、P<0.01)。(2)与单药组相比,联合用药组中SW1990细胞增殖率显著降低,克隆形成率显著降低,凋亡率显著升高(P<0.05),细胞中的Bcl-2蛋白表达水平显著降低。(3)与单药组相比,联合用药组细胞迁移率显著降低,细胞中的EMT相关蛋白E-钙黏蛋白表达显著升高,波形蛋白表达显著降低。结论阿司匹林有抑制胰腺癌细胞增殖的作用,并且能增强细胞对吉西他滨的敏感性,同时逆转胰腺癌细胞EMT。

Objective To investigate the effects of aspirin on the epithelial mesenchymal transition and gemcitabine resistance of human pancreatic cancer cells.Methods Three kinds of human pancreatic cancer cell lines SW1990,PATU-8988 and PANC-1were treated with aspirin in different concentrations of 0,1,2,5and 10mmol/L,respectively.The SW1990 cells were treated with combined aspirin 2mmol/L and gemcitabine 0.05mg/L.The inhibition of cell proliferation was detected by MTT assay.Clonogenic assay was used to detect the proliferation ability.Wound healing assay was used to observe the migration capability.Flow cytometry was used to detect the apoptosis and Western blot was used to detect the expressions of the proteins related to EMT and apoptosis.Results Compared with the blank control group,the low dose（1,2mmol/L）of aspirin inhibited the proliferation of PATU-8988 and PANC-1（P0.05）,the high dose（5,10mmol/L）of aspirin inhibited the proliferation of the three kinds of human pancreatic cancer cell lines（P0.05）.Compared with using aspirin or gemcitabine alone,combined use of aspirin and gemcitabine could reduce the SW1990 cell proliferation rate and clone formation rate,increase the apoptotic rate（P0.05）,decrease the expression of Bcl-2.Compared with using aspirin or gemcitabine alone,combined use of aspirin and gemcitabine could reduce the cell migration rate,increase the expression of E-cadherin,reduce the expression of Vimentin,which were related with the epithelial mesenchymal transition.Conclusion Aspirin may inhibit the proliferation of pancreatic cancer cell,enhance its sensitivity to gemcitabine,and reverse the epithelial mesenchymal transition of the pancreatic cancer cells.