Wang J F, Guo Z
Department of Colorectal Oncology, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China.
Department of Interventional Therapy, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China.
Zhonghua Nei Ke Za Zhi. 2025 Jul 1;64(7):670-674. doi: 10.3760/cma.j.cn112138-20241115-00757.
To assess the anti-tumor immune response after combined cryoablation and granulocyte-macrophage colony-stimulating factor (GM-CSF) treatment. A mouse model of colorectal liver metastases (CLM) was established using 80 BALB/c mice. All tumor-bearing mice were divided into a control group (=20), a GM-CSF group (=20), a cryoablation group (=20), and a cryoablation+GM-CSF group (=20). Cryoablation was applied using a 1.7-mm diameter cryosurgery needle from an argon-helium cryosurgery system. Tumor size and survival time were observed. Changes in the number of dendritic cells (DCs) in tumors were assessed by immunohistochemistry. Serum cytokine (IFN-γ, IL-4) levels were analyzed by enzyme-linked immunosorbent assay (ELISA), and the Th1/Th2 ratio was estimated from the IFN-γ/IL-4 ratio. One-way analysis of variance was used for data analysis. Our findings showed that combining cryoablation and GM-CSF synergistically increased infiltrating DC numbers. Among the four groups, cryoablation+GM-CSF group exhibited the highest DC count post-treatment (control group: 1.4±0.5/HPF, GM-CSF group: 15.3±1.8/HPF, cryoablation group: 9.6±1.5/HPF, cryoablation+GM-CSF group:17.4±1.3/HPF; =138.60,<0.05). Cryoablation, alone or combined with GM-CSF, elevated serum IFN-γ and reduced serum IL-4. IFN-γ levels of cryoablation group increased from 45.6±2.4 pg/ml to 57.8±3.3 pg/ml,=170.09,<0.05,and IFN-γ levels of cryoablation+GM-CSF group increased from 45.6±2.4 pg/ml to 86.3±1.9 pg/ml,=664.31,<0.05;and IL-4 levels of cryoablation group decreased from 7.21±0.63 pg/ml to 6.35±0.25 pg/ml,=5.77,<0.05,IL-4 levels of cryoablation+GM-CSF group decreased from 7.21±0.63 pg/ml to 6.42±0.21 pg/ml,=10.93,<0.05. Conversely, GM-CSF alone decreased serum IFN-γ and increased serum IL-4. Notably, combined cryoablation and GM-CSF significantly boosted Th1/Th2 ratios (cryoablation+GM-CSF group: from 6.32±0.45 to 13.43±0.64; =379.95,<0.05). Combined cryoablation with GM-CSF treatment exerted a promising anti-tumor immune effect in the treatment of colorectal cancer liver metastases. This approach increased the number of DCs in the tumor microenvironment and improved the anti-tumor immunologic response in tumor-bearing mice.
评估冷冻消融联合粒细胞巨噬细胞集落刺激因子（GM-CSF）治疗后的抗肿瘤免疫反应。使用80只BALB/c小鼠建立结直肠癌肝转移（CLM）小鼠模型。所有荷瘤小鼠分为对照组（n = 20）、GM-CSF组（n = 20）、冷冻消融组（n = 20）和冷冻消融+GM-CSF组（n = 20）。使用氩氦冷冻手术系统的1.7毫米直径冷冻手术针进行冷冻消融。观察肿瘤大小和生存时间。通过免疫组织化学评估肿瘤中树突状细胞（DCs）数量的变化。通过酶联免疫吸附测定（ELISA）分析血清细胞因子（IFN-γ、IL-4）水平，并根据IFN-γ/IL-4比值估算Th1/Th2比值。采用单因素方差分析进行数据分析。我们的研究结果表明，冷冻消融与GM-CSF联合使用可协同增加浸润性DC数量。在四组中，冷冻消融+GM-CSF组治疗后DC计数最高（对照组：1.4±0.5/HPF，GM-CSF组：15.3±1.8/HPF，冷冻消融组：9.6±1.5/HPF，冷冻消融+GM-CSF组：17.4±1.3/HPF；F = 138.60，P<0.05）。单独冷冻消融或与GM-CSF联合使用，均可提高血清IFN-γ水平并降低血清IL-4水平。冷冻消融组IFN-γ水平从45.6±2.4 pg/ml升高至57.8±3.3 pg/ml，F = 170.09，P<0.05，冷冻消融+GM-CSF组IFN-γ水平从45.6±2.4 pg/ml升高至86.3±1.9 pg/ml，F = 664.31，P<0.05；冷冻消融组IL-4水平从7.21±0.63 pg/ml降低至6.35±0.25 pg/ml，F = 5.77，P<0.05，冷冻消融+GM-CSF组IL-4水平从7.21±0.63 pg/ml降低至6.42±0.21 pg/ml，F = 10.93，P<0.05。相反，单独使用GM-CSF会降低血清IFN-γ水平并升高血清IL-4水平。值得注意的是，冷冻消融与GM-CSF联合使用可显著提高Th1/Th2比值（冷冻消融+GM-CSF组：从6.32±0.45提高至13.43±0.64；F = 379.95，P<0.05）。冷冻消融与GM-CSF联合治疗在结直肠癌肝转移治疗中具有良好的抗肿瘤免疫效果。这种方法增加了肿瘤微环境中DC的数量，并改善了荷瘤小鼠的抗肿瘤免疫反应。
