高良姜素调节Notch-1/Hes信号通路对胰腺癌细胞恶性生物学行为的影响

Effect of galangin on malignant biological behavior of pancreatic cancer cells by regulating Notch-1/Hes signaling pathway

目的:探讨高良姜素(Gal)调节Notch-1/Hes信号通路对胰腺癌细胞恶性生物学行为的影响。方法:将人胰腺癌PCNA-1细胞分为空白组、Gal低剂量组、Gal中剂量组、Gal高剂量组、Jagged1组、Gal高剂量+Jagged1组,EdU染色和CCK-8、划痕实验、Transwell实验分别检测PCNA-1细胞增殖、迁移、侵袭;qRT-PCR检测PCNA-1细胞中细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶(MMP)-9、MMP-2 mRNA表达;裸鼠体内移植瘤实验检测裸鼠体内肿瘤质量与体积;Western blot检测PCNA-1细胞及肿瘤组织中Notch-1、Hes1蛋白表达。结果:与空白组相比,Gal低剂量组、Gal中剂量组、Gal高剂量组PCNA-1细胞EdU阳性细胞率、OD 450值、划痕愈合率、细胞侵袭数目、Cyclin D1、MMP-9、MMP-2 mRNA表达、PCNA-1细胞中Notch-1、Hes1蛋白表达降低,且呈剂量依赖性,Jagged1组对应指标变化趋势与上述相反(P<0.05);与Gal高剂量组相比,Gal高剂量+Jagged1组PCNA-1细胞EdU阳性细胞率、OD 450值、划痕愈合率、细胞侵袭数目、Cyclin D1、MMP-9、MMP-2 mRNA表达、PCNA-1细胞中Notch-1、Hes1蛋白表达升高(P<0.05)。与裸鼠-空白组相比,裸鼠-Gal低剂量组、裸鼠-Gal中剂量组、裸鼠-Gal高剂量组裸鼠肿瘤质量与体积、Notch-1、Hes1蛋白表达降低,且呈剂量依赖性(P<0.05),裸鼠-Jagged1组对应指标变化趋势与上述相反(P<0.05);与裸鼠-Gal高剂量组相比,裸鼠-Gal高剂量+Jagged1组裸鼠肿瘤质量与体积、Notch-1、Hes1蛋白表达升高(P<0.05)。结论:Gal抑制PCNA-1细胞增殖、迁移、侵袭及裸鼠体内肿瘤的生长,可能与抑制Notch-1/Hes通路有关。

Objective:To investigate the effect of galangin(Gal)on the malignant biological behavior of pancreatic cancer cells by regulating Notch-1/Hes signaling pathway.Methods:Human pancreatic cancer PCNA-1 cells were grouped into blank group,Gal low-dose group,Gal medium-dose group,Gal high-dose group,Jagged1 group,and Gal high-dose+Jagged1 group.EdU staining,CCK-8,scratch assay,and Transwell assay were applied to detect the proliferation,migration,and invasion of PCNA-1 cells,respectively.qRT-PCR was applied to detect the expression of Cyclin D1,matrix metalloproteinase-9,and MMP-2 mRNA in PCNA-1 cells.Nude mouse in vivo tumor transplantation experiment was applied to detect the mass and volume of tumors in nude mice.Western blot was applied to detect the expression of Notch-1 and Hes1 proteins in PCNA-1 cells and tumor tissues.Results:Compared with the blank group,the EdU positive cell rate,OD 450 value,scratch healing rate,cell invasion number,Cyclin D1,MMP-9,MMP-2 mRNA expression,Notch-1,Hes1 protein expression in PCNA-1 cells were reduced in the Gal low-dose group,Gal medium-dose group,and Gal high-dose group,and were dose-dependent,the change trend of corresponding indicators in Jagged1 group was opposite to the above(P<0.05).Compared with the Gal high-dose group,the EdU positive cell rate,OD 450 value,scratch healing rate,number of cell invasions,Cyclin D1,MMP-9,MMP-2 mRNA expression,and Notch-1 and Hes1 protein expression in PCNA-1 cells in Gal high-dose+Jagged1 group increased(P<0.05).Compared with the nude mice-blank group,the tumor mass and volume,and the expression of Notch-1 and Hes1 proteins in the nude mouse Gal low-dose group,the nude mouse Gal medium-dose group,and the nude mouse Gal high-dose group were reduced,in a dose-dependent manner(P<0.05),the change trends of corresponding indicators in the nude mice-Jagged1 group were opposite to the above(P<0.05).Compared with the nude mice high-dose group,the tumor mass and volume,and the expression of Notch-1 and Hes1 proteins in the nude mice Gal high-dose+Jagged1 group increased(P<0.05).Conclusion:Gal inhibits the proliferation,migration,invasion,and tumor growth of PCNA-1 cells in nude mice,which may be related to the inhibition of the Notch-1/Hes pathway.