牛蒡子苷元调控Notch/Hes-1信号通路对口腔鳞状细胞癌HSC-3细胞增殖、凋亡和侵袭的影响

Effects of arctigenin on the proliferation,apoptosis,and invasion of oral squamous cell carcinoma HSC-3 cells by regulating the Notch/Hes-1 signaling pathway

目的:探究牛蒡子苷元(ARC)通过调控Notch/Hes-1信号通路对口腔鳞状细胞癌(OSCC)HSC-3细胞增殖、凋亡和侵袭的影响及其机制。方法:使用不同质量浓度的ARC处理人HSC-3细胞,CCK-8法检测ARC对细胞增殖活力的影响,以选择适宜的药物浓度。将HSC-3细胞分为对照组、ARC-L组(10 mg/L ARC)、ARC-M组(20 mg/L ARC)、ARC-H组(40 mg/L ARC)和ARC-H+Jagged1/FC组(40 mg/L ARC+1.2μg/mL Jagged1/FC)。采用EdU法检测细胞增殖能力,划痕愈合实验、Transwell实验和流式细胞术分别检测细胞的迁移、侵袭能力及细胞周期和细胞凋亡率,WB法检测增殖(c-Myc、cyclin D1)、凋亡(BAX、Bcl-2、survivin)、EMT(E-cadherin、vimentin、Snail)及Notch/Hes-1通路(Notch 1、Hes-1、NICD)相关蛋白的表达水平。结果:与0 mg/L相比,10~80 mg/L的ARC均能显著降低HSC-3细胞增殖活力(均P<0.05)。与对照组相比,ARC-L组、ARC-M组和ARC-H组HSC-3细胞EdU阳性率、划痕愈合率、侵袭细胞数、S期和G2/M期细胞占比及c-Myc、cyclin D1、Bcl-2、survivin、vimentin、Snail、Notch 1、Hes-1和NICD蛋白表达均显著降低(均P<0.05),细胞凋亡率、G0/G1期细胞占比及BAX、E-cadherin的蛋白表达均显著升高(均P<0.05),且呈浓度梯度依赖性。同时使用Notch激动剂Jagged1/FC,则可部分逆转ARC对HSC-3细胞增殖、迁移、侵袭、凋亡及相关蛋白表达的作用(均P<0.05)。结论:ARC可能通过抑制Notch/Hes-1信号通路抑制OSCC细胞HSC-3增殖和侵袭并促进细胞凋亡。

Objective:To investigate the effects of arctigenin(ARC)on the proliferation,apoptosis and invasion of oral squamous cell carcinoma(OSCC)HSC-3 cells by regulating the Notch/Hes-1 signaling pathway and its mechanism.Methods:Human HSC-3 cells were treated with different mass concentrations of ARC,and the effect of ARC on cell proliferation was detected by CCK-8 method to select the appropriate drug concentration.HSC-3 cells were divided into the control group,the ARC-L group(10 mg/L ARC),the ARC-M group(20 mg/L ARC),the ARC-H group(40 mg/L ARC),and the ARC-H+Jagged1/FC group(40 mg/L ARC+1.2μg/mL Jagged1/FC).Cell proliferation ability was detected by EdU assay,and cell migration and invasion abilities,cell cycle and apoptosis rate were detected by scratch healing assay,Transwell assay and flow cytometry,respectively.The expression levels of proliferation(c-Myc,cyclin D1),apoptosis(BAX,Bcl-2,survivin),EMT(E-cadherin,vimentin,Snail)and Notch/Hes-1 pathway(Notch1,Hes-1,NICD)related proteins were detected by Western blot.Results:Compared with 0 mg/L,10-80 mg/L of ARC significantly decreased the proliferation vitality of HSC-3 cells(all P<0.05).Compared with the control group,the EdU positive rate,scratch healing rate,invasion cell number,proportions of S phase and G2/M phase cells,and the expressions of c-Myc,cyclin D1,Bcl-2,survivin,vimentin,Snail,Notch 1,Hes-1,and NICD proteins of HSC-3 cells decreased significantly in the ARC-L,ARC-M,and ARC-H groups(all P<0.05);the apoptosis rate,the proportion of G0/G1 phase cells,and the expressions of BAX and E-cadherin proteins increased significantly(all P<0.05),and were concentration gradient dependent.Simultaneous use of Notch agonist Jagged1/FC partially reversed the effects of ARC on the proliferation,migration,invasion,apoptosis and the expressions of related proteins of HSC-3 cells(all P<0.05).Conclusion:ARC may inhibit the proliferation and invasion of OSCC HSC-3 cells and promote cell apoptosis by inhibiting the Notch/Hes-1 signaling pathway.