[摘 要] 目的：探究银杏内酯B（GKB）调控蛋白激酶R样内质网激酶（PERK）/转录激活子4（ATF4）/C/EBP同源蛋白（CHOP） 信号通路对肝癌细胞增殖、迁移、凋亡和上皮间质转化（EMT）的影响。方法：将人肝癌细胞MHCC-97H随机分为对照组、GKB 组、GSK2656157（PERK抑制剂）组和GKB + GSK2656157组，以GKB和GSK2656157分别干预后，采用MTT法和EdU染色检测 各组细胞的增殖活性及增殖率，划痕愈合实验、流式细胞术分别检测各组细胞的迁移及凋亡水平，WB法检测各组细胞中EMT和 PERK/ATF4/CHOP信号通路相关蛋白的表达水平。构建MHCC-97H细胞裸鼠移植瘤模型，以同法分组及药物干预后测定各组 移植瘤体积，采用免疫组化、TUNEL染色分别检测各组肿瘤细胞增殖、凋亡水平，WB法检测各组移植瘤组织中EMT和PERK/ ATF4/CHOP信号通路相关蛋白的表达水平。结果：与对照组比较，GKB组细胞活性、增殖率、迁移率、移植瘤体积、Ki-67阳性细 胞率、MMP2、N-cadherin与MMP9蛋白表达均显著降低（均 P < 0.05），细胞凋亡率、TUNEL 阳性细胞率、p-PERK/PERK与 E-cadherin、ATF4、CHOP蛋白表达均显著升高（均P < 0.05）；GSK2656157组各指标变化与GKB组相反（均P < 0.05）。与GKB组 比较，GKB + GSK2656157组细胞活性、增殖率、迁移率、移植瘤体积、Ki-67阳性细胞率、MMP2、N-cadherin与MMP9蛋白表达均 显著升高（均P < 0.05），细胞凋亡率、TUNEL 阳性细胞率、p-PERK/PERK 与 E-cadherin、ATF4、CHOP 蛋白表达均显著降低（均 P < 0.05）。结论：GKB可通过激活PERK/ATF4/CHOP信号通路抑制肝癌MHCC-97H细胞增殖、迁移和EMT并促进其凋亡。

[Abstract] Objective: To investigate the effects of ginkgolide B (GKB) on the proliferation, migration, apoptosis, and epithelialmesenchymal transition (EMT) of liver cancer cells by regulating the protein kinase R-like endoplasmic reticulum kinase (PERK)/ activating transcription factor 4 (ATF4)/C/EBP homologous protein (CHOP) signaling pathway. Methods: Human liver cancer cells MHCC-97H were randomly assigned into the control group, the GKB group, the GSK2656157 (PERK inhibitors) group, and the GKB + GSK2656157 group. After intervention with GKB and PERK inhibitors GSK2656157 on different groups, MTT assay and EdU staining were used to detect the proliferation activity and proliferation rate of cells in each group. Scratch assay and flow cytometry were used to detect the migration and apoptosis of cells in each group, respectively. Western blotting (WB) was used to detect the expression levels of EMT and PERK/ATF4/CHOP signaling pathway related proteins in each group. The MHCC-97H nude mouse transplant tumor model was constructed, and the tumor volumes of each group were measured after grouping and drug intervention. Immunohistochemistry and TUNEL staining were used to detect the proliferation and apoptosis of tumor cells in each group. In addition, WB was used to detect the expression levels of EMT and PERK/ATF4/CHOP signaling pathway related proteins in transplant tumor tissues of each group. Results: Compared with those in the control group, the cell activity, proliferation rate, migration rate, tumor volume, Ki-67 positive cell ratio, and relative expressions of MMP2, N-cadherin, and MMP9 proteins decreased significantly in the GKB group (all P < 0.05), while the apoptosis rate, TUNEL positive cell ratio, and the expressions of p-PERK/PERK, E-cadherin, ATF4, and CHOP proteins increased significantly (all P < 0.05). The changes in various indicators in the GSK2656157 group were the opposite of those in the GKB group (all P < 0.05). Compared with those in the GKB group, the cell activity, proliferation rate, migration rate, tumor volume, Ki-67 positive cell ratio, and expressions of MMP2, N-cadherin, and MMP9 proteins increased significantly in the GKB + GSK2656157 group (all P < 0.05), while the apoptosis rate, TUNEL positive cell ratio, and the expressions of p-PERK/PERK, E-cadherin, ATF4, and CHOP proteins reduced significantly (all P < 0.05). Conclusion: GKB can inhibit the proliferation, migration, and EMT of liver cancer MHCC-97H cells and promote their apoptosis by activating the PERK/ATF4/CHOP signaling pathway.

[基金项目] 南通市自然科学基金和社会民生计划项目（MSZ2023077）