四氢生物蝶呤对胰腺癌细胞增殖、迁移及凋亡的影响

Effects of BH4 on proliferation,migration and apoptosis of pancreatic cancer cells

目的探究四氢生物蝶呤(tetrahydrobiopterin,BH4)对胰腺癌细胞生物学行为的影响及可能机制。方法细胞分为对照组与实验组,实验组根据BH4浓度(2、4、8、12µM)分为4组,采用CCK-8、Hoechst及划痕实验检测细胞的增殖、凋亡和迁移能力,RT-qPCR检测细胞中Notch1 mRNA表达情况,Western blotting检测细胞中E-Cadherin与N-cadherin表达情况。结果与对照组相比较,随着BH4浓度的增加,HPAC与BxPC-3细胞的活性与迁移能力逐渐下降,细胞凋亡率逐渐增高,且8µM、12µM浓度的BH4作用有显著意义(P<0.01,P<0.05);与对照组相比较,12µM BH4组细胞中Notch1 mRNA显著降低(P<0.05),E-cadherin蛋白表达水平升高(P<0.01,P<0.05),N-cadherin蛋白表达水平降低(P<0.05)。结论BH4可抑制胰腺癌细胞的增殖和迁移,促进其凋亡,其机制可能与抑制上皮细胞-间充质转化(epithelial-mesenchymal transition,EMT)有关,而Notch1在EMT发生过程中改变,提示Notch1信号通路可能参与该过程的调控。

Objective To investigate the effect of tetrahydrobiopterin(BH4)on the biological behavior of pancreatic cancer cells and its possible mechanism.Methods The cells were divided into a control group and an experimental group.The experimental group was divided into 4 subgroups according to the concentration of BH4(2µM,4µM,8µM,12µM BH4 groups).The ability of cell proliferation,apoptosis and migration were detected by CCK-8 assay,Hoechst assay and scratch assay.The expression of Notch1 mRNA was detected by RT-qPCR and the expressions of E-cadherin and N-cadherin were detected by Western blotting.Results Compared with the control group,with the increasing of BH4 concentration,the activity and migration ability of HPAC and Bxpc-3 cell decreased and the apoptosis rate increased,and the effect of BH4 was significant at 8µM and 12µM(P<0.01,P<0.05).Compared with the control group,Notch1 mRNA was significantly decreased in experimental group(BH412µM)(P<0.05),the expression level of E-cadherin increased(P<0.01,P<0.05),while that of N-cadherin decreased(P<0.05).Conclusions BH4 can inhibit the proliferation,migration and promote the apoptosis of pancreatic cancer cells.The mechanism may be related to the inhibition of epithelial-mesenchymal transition(EMT)process,while Notch1 changes during EMT.It suggests that the Notch1 signaling pathway may be involved in the regulation of this process.