[摘 要] 目的：探究肺癌磷酸甘油酸变位酶1（PGAM1）对肺癌LLC细胞增殖和迁移的调控作用，以及PGAM1对肿瘤微环境 中CD8+ T细胞功能和浸润的影响。方法：将shPGAM1及shNC慢病毒感染LLC细胞，筛选稳定细胞，分别命名为shPGAM1组 和NC组，WB和qPCR法检测两组细胞中PGAM1蛋白和mRNA的表达水平。CCK-8法和实时细胞分析仪检测敲低PGAM1对 LLC细胞增殖和迁移的影响；流式细胞术检测与两组细胞分别体外共培养后CD8+ T细胞功能性受体（TIM-3、PD-1、 GrzmB、Ki67）表达的变化。构建小鼠肺癌LLC细胞皮下移植瘤模型，监测PGAM1表达对肿瘤生长的影响，通过流式细胞术检测 敲低PGAM1对肿瘤微环境中CD8+ T细胞浸润的影响。结果：shPGAM1组细胞中PGAM1蛋白和mRNA表达水平较NC组均显 著降低。敲低PGAM1可降低LLC细胞的增殖和迁移能力（Ρ < 0.000 1或Ρ < 0.05）。与shPGAM1组细胞共培养后，CD8+ T细 胞的耗竭标志物（TIM-3、PD-1）表达均降低（Ρ < 0.000 1或Ρ < 0.01）。敲低PGAM1后肿瘤生长受到显著抑制，且肿瘤组织中 CD8+ T细胞浸润增加（Ρ

[Abstract] Objective: To investigate the regulatory role of phosphoglycerate mutase 1 (PGAM1) in lung cancer LLC cell proliferation and migration, and to explore its impact on the function and infiltration of CD8? T cells within the tumor microenvironment. Methods: LLC cells were infected with shPGAM1 and shNC lentiviruses, and stable cell lines were established and designated as the shPGAM1 group and the NC group, respectively. WB and qPCR were used to assess the protein and mRNA expression levels of PGAM1 in both groups. Cell proliferation and migration after PGAM1 knockdown were assessed using the CCK-8 assay and real-time cell analyzer. Flow cytometry was employed to detect the expression of CD8? T-cell functional receptors (TIM-3, PD-1, GrzmB, Ki67) after co culture with cells from each group in vitro. A subcutaneous LLC xenograft model was established in mice to examine the effect of PGAM1 on tumor growth, and flow cytometry was used to evaluate the impact of PGAM1 knockdown on CD8? T cell infiltration in the tumor microenvironment. Results: The protein and mRNA expression levels of PGAM1 were significantly reduced in the shPGAM1 group compared with the NC group. Knockdown of PGAM1 significantly reduced the proliferation and migration of LLC cells (P < 0.000 1 or P < 0.05). After co-culture with shPGAM1 cells, the expression of exhaustion markers (TIM-3, PD-1) on CD8? T cells decreased notably (P < 0.000 1 or P < 0.01). Knockdown of PGAM1 significantly inhibited tumor growth and increased CD8? T cell infiltration in tumor tissues (P < 0.000 1). Conclusion: Targeted knockdown of PGAM1 inhibits tumor cell proliferation and migration, reduces the expression of exhaustion markers on CD8? T cells, and enhances T cell infiltration within tumors, thereby exerting dual effects on tumor growth and antitumor immunity.

[基金项目] 国家自然科学基金（No.82373283）