摘要
目的:探讨高良姜素(Gal)调控AMPK/mTOR/ULK1信号通路对胃癌细胞凋亡和自噬的影响及其机制。方法:将胃癌NCI-N87细胞分为对照组、多索吗啡(DM)组、Gal低剂量(Gal-L)组、Gal高剂量(Gal-H)组、Gal-H+DM组。采用MTT法、流式细胞术、划痕愈合实验和Transwell实验分别检测各组细胞的增殖、凋亡、迁移和侵袭能力,WB法检测PCNA、C-caspase-3、免疫逃逸相关蛋白(B7H1)、EMT和AMPK/mTOR/ULK1信号通路蛋白的表达水平。建立裸鼠NCI-N87细胞移植瘤模型,观察Gal和5-FU对移植瘤的抑制效果。结果:与对照组比较,DM组NCI-N87细胞增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著升高(均P<0.05),细胞凋亡率、C-caspase-3、E-cadherin、LC3II/LC3I、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著降低(均P<0.05);Gal-L组和Gal-H组NCI-N87细胞的增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著降低(均P<0.05),细胞凋亡率、C-caspase-3、E-cadherin、LC3II/LC3I、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著升高(均P<0.05);DM可部分逆转Gal对NCI-N87细胞恶性生物学行为的抑制作用(P<0.05);与对照组比较,Gal组和5-FU组裸鼠移植瘤体积和质量均显著降低,肿瘤组织细胞凋亡率显著升高(P<0.05)。结论:Gal可促进胃癌NCI-N87细胞自噬和凋亡,抑制其增殖、迁移和侵袭,可能与激活AMPK/mTOR/ULK1信号通路有关。
Objective:To investigate the effects and mechanisms of galangin(Gal)on the apoptosis and autophagy of gastric cancer NCI-N87 cells through regulating the AMPK/mTOR/ULK1 signaling pathway.Methods:Gastric cancer NCI-N87 cells were assigned into the control group,the dorsomorphin(DM,AMPK inhibitor)group,the Gal low-dose(Gal-L)group,the Gal high-dose(Gal-H)group,and the Gal-H+DM group.Cell proliferation,apoptosis,migration,and invasion abilities were detected using MTT assay,flow cytometry,wound healing assay,and Transwell assay,respectively.Western blotting was used to detect the expression levels of PCNA,cleaved caspase-3(C-caspase-3),immune evasion-related protein(B7H1),EMT,and AMPK/mTOR/ULK1 signaling pathway proteins.A NCI-N87 cell xenograft tumor model was established in nude mice to observe the inhibitory effects of Gal and 5-FU on the growth of transplant tumors.Results:Compared with the control group,the DM group exhibited significantly increased proliferation activity,scratch healing rate,number of invasive cells,and protein expression levels of N-cadherin,vimentin,PCNA,B7H1,p62,and p-mTOR/mTOR in NCI-N87 cells(all P<0.05),along with significantly decreased apoptosis rate and protein expression levels of C-caspase-3,E-cadherin,LC3Ⅱ/LC3Ⅰ,p-AMPK/AMPK,and p-ULK1/ULK1(all P<0.05).In contrast,both the Gal-L and Gal-H groups showed significantly decreased proliferation activity,scratch healing rate,number of invasive cells,and protein expression levels of N-cadherin,vimentin,PCNA,B7H1,p62,and p-mTOR/mTOR in NCI-N87 cells(all P<0.05),while displaying significantly increased apoptosis rate and protein expression levels of C-caspase-3,E-cadherin,LC3Ⅱ/LC3Ⅰ,p-AMPK/AMPK,and p-ULK1/ULK1(all P<0.05).DM could partially reverse the inhibitory effect of Gal on the malignant biological behaviors of NCI-N87 cells(P<0.05).Compared with the control group,both the Gal group and the 5-FU group exhibited significant reductions in tumor volume and mass,as well as a significant increase in the apoptosis rate of tumor tissue cells in nude mice(P<0.05).Conclusion:Gal can promote the autophagy and apoptosis in gastric cancer NCI-N87 cells,inhibit their proliferation,migration,and invasion,which may be related to the activation of the AMPK/mTOR/ULK1 signaling pathway.
作者
郭芳
陈巍
刘蒙
邹艳丽
田霞
GUO Fang;CHEN Wei;LIU Meng;ZOU Yanli;TIAN Xia(Department of Gastroenterology,the Third Hospital of Wuhan,Wuhan 430000,Hubei,China)
出处
《中国肿瘤生物治疗杂志》
北大核心
2026年第1期59-65,共7页
Chinese Journal of Cancer Biotherapy
基金
武汉市医学科学研究项目(WX23Z20)。
