[摘 要] 目的：探究包被蛋白复合体β1亚基（COPB1）在食管鳞状细胞癌（ESCC）中的表达，及其对ESCC细胞恶性生物学行 为的影响、作用机制及临床意义。方法：采用2014～2018年间在河北医科大学第四医院生物样本库中82例ESCC组织及癌旁组 织，常规培养正常食管鳞状上皮细胞HEEC和食管癌细胞KYSE-150、KYSE-170、Eca109、TE1、KYSE-30、KYSE-450，用转染试剂 将pcDNA3.1-vecto（r 空载体）、pcDNA3.1-COPB1载体，si-NC和si-COPB1转染至KYSE-150、TE1细胞中，记为NC、COPB1-OE、 si-NC和si-COPB1组。用数据库数据分析COPB1 mRNA在泛癌组织中的表达及其表达与免疫细胞浸润的关系，qPCR法检测 ESCC组织和细胞中COPB1、PIK3CB、CD68、CD163、CD206、ARG1、IL-10 mRNA水平表达情况，WB法检测ESCC组织和各组细 胞中的COPB1、PI3K、CD68、CD163、CD206、p-AKT蛋白表达，克隆形成实验和MTS实验检测各组细胞的增殖能力，划痕愈合实 验和Transwell实验检测各组细胞的迁移和侵袭能力，免疫组织化学染色（IHC）法检测ESCC组织中COPB1和CD206蛋白表达。 以人单核细胞白血病细胞（THP-1）构建巨噬细胞模型，用佛波酯（PMA）和IL-3和IL-4和ESCC细胞上清液诱导巨噬细胞转型，用 qPCR和WB法检测CD68和CD206m RNA和蛋白的表达。结果：COPB1在泛癌组织和ESCC组织中均呈高表达且与淋巴结转 移和TNM分期有关联（均P < 0.01），COPB1高表达的ESCC患者总生存期短（P < 0.05），COPB1是潜在的ESCC的诊断标志物。 COPB1在KYSE-150和TE1细胞中也呈高表达（均P < 0.05），过表达或敲减COPB1可明显抑制或促进KYSE-150和TE1细胞的 增殖能力、迁移和侵袭能力（均 P < 0.05）。COPB1 表达变化诱导的差异表达基因主要富集于 PI3K/AKT 通路（均 P < 0.001）, COPB1可促进PI3K/AKT通路的活化（P < 0.05），COPB1 高表达可导致 M2 型巨噬细胞浸润增加（P < 0.05），COPB1高表达 促进TAM/M2极化（P < 0.05）。结论：COPB1在ESCC组织中呈高表达，其可激活PI3K/AKT通路及调控肿瘤免疫微环境促进 ESCC发生发展，COPB1有望成为ESCC诊断和预后的生物标志物及治疗靶点。

[Abstract] Objective: To investigate the expression of coatomer protein complex subunit beta 1 (COPB1) in esophageal squamous cell carcinoma (ESCC) and its impact on the malignant biological behavior of ESCC cells, as well as the underlying mechanism of action and its clinical significance. Methods: A total of 82 pairs of ESCC tissues and adjacent non-cancerous were collected from the biobank of the Fourth Hospital of Hebei Medical University during 2014 and 2018. Normal esophageal squamous cells (HEEC) and ESCC cell lines (KYSE-150, KYSE-170, Eca109, TE1, KYSE-30, KYSE-450) were routinely cultured. The pcDNA3.1-vector (empty vector), pcDNA3.1-COPB1 vector, si-NC, and si-COPB1 were transfected into KYSE-150 and TE1 cells using transfection reagents. The resulting groups were designated as the NC, COPB1-OE, si-NC, and si-COPB1 groups, respectively. A comprehensive database analysis was conducted to assess the mRNA expression of COPB1 in pan-cancer tissues and its correlation with immune cell infiltration. Quantitative real-time polymerase chain reaction (qPCR) was utilized to assess the mRNA expression of COPB1, PIK3CB, CD68, CD163, CD206, ARG1, and IL-10 in ESCC tissues and cells. Western blotting (WB) analysis was performed to detect COPB1, PI3K, CD68, CD163, CD206 and p-AKT protein expression in ESCC tissues and various cell groups. Colony formation and MTS assays were conducted to assess cell proliferation, while wound healing assay and Transwell assay were used to determine cell migration and invasion. Immunohistochemistry (IHC) was employed to detect the protein expression of COPB1 and CD206 in ESCC tissues. A macrophage model was established using human monocytic leukemia cells (THP-1). Macrophage polarization was induced with phorbol myristate acetate (PMA), interleukin-3 (IL-3), IL-4, and ESCC cell supernatant. WB and qPCR were performed to detect the protein and mRNA expression of CD68 and CD206, respectively. Results: COPB1 was highly expressed in both pan-cancer and ESCC tissues, and its expression was associated with lymph node metastasis and TNM staging (both P < 0.01). ESCC patients with high COPB1 expression exhibited shorter overall survival (P < 0.05), making COPB1 a potential diagnostic biomarker for ESCC. COPB1 was also highly expressed in KYSE-150 and TE1 cells (both P < 0.05). Overexpression or knockdown of COPB1 significantly inhibited or promoted the proliferation, migration and invasion capabilities of KYSE-150 and TE1 cells (both P < 0.05). Differentially expressed genes induced by COPB1 expression changes were primarily enriched in the PI3K/AKT pathway (both P < 0.001), and COPB1 activated the PI3K/AKT pathway (P < 0.05). Additionally, high COPB1 expression increased M2 macrophage infiltration (P < 0.05) and promoted TAM/M2 polarization (P < 0.05). Conclusion: COPB1 is highly expressed in ESCC tissues and can activate the PI3K/AKT pathway and modulates the tumor immune microenvironment, thereby promoting ESCC occurrence and development. COPB1 holds promise as a diagnostic and prognostic biomarker, as well as a therapeutic target for ESCC.

[基金项目] 河北省重点研发计划（No. 22377730D；No. 22377721D）；河北省自然科学基金项目（No. H2025206589）；河北医科 大学第四医院科研创新团队支持计划（No. 2023C12）；政府资助临床医学优秀人才培养项目（No. ZF2024112）