[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。

[Abstract] 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.

[基金项目] 武汉市医学科学研究项目（WX23Z20）