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、12 μM）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8 μM、12 μM浓度的BH4作用有显著意义（P<0.01，P<0.05）；与对照组相比较，12 μM 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。
Abstract
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 (2 μM, 4 μM, 8 μM, 12 μM 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 8 μM and 12 μM (P<0.01，P<0.05). Compared with the control group, Notch1 mRNA was significantly decreased in experimental group (BH4 12 μM) (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.
关键词
四氢生物蝶呤 /
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胰腺癌 /
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上皮间质转化 /
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Notch信号通路
Key words
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BH4 /
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Pancreatic cancer /
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Epithelial-mesenchymal transition /
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Notch signaling pathway
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基金
国家自然科学基金青年基金（81502313）
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