[Experimental study of anti-tumor immunologic response functions after combined treatment of cryoablation with GM-CSF for colorectal cancer liver metastases]

Zhonghua Nei Ke Za Zhi. 2025 Jul 1;64(7):670-674. doi: 10.3760/cma.j.cn112138-20241115-00757.
[Article in Chinese]

Abstract

Objective: To assess the anti-tumor immune response after combined cryoablation and granulocyte-macrophage colony-stimulating factor (GM-CSF) treatment. Methods: 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 (n=20), a GM-CSF group (n=20), a cryoablation group (n=20), and a cryoablation+GM-CSF group (n=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. Results: 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; F=138.60,P<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,F=170.09,P<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,F=664.31,P<0.05;and IL-4 levels of cryoablation group decreased from 7.21±0.63 pg/ml to 6.35±0.25 pg/ml,F=5.77,P<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,F=10.93,P<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; F=379.95,P<0.05). Conclusion: 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.

目的: 探讨冷冻消融联合粒细胞-巨噬细胞集落刺激因子(GM-CSF)治疗后机体抗肿瘤免疫反应。 方法: 建立结直肠癌肝转移(CLM)小鼠模型。随机分为对照组(n=20)、冷冻消融组(n=20)、GM-CSF组(n=20)、冷冻消融+GM-CSF联合组(n=20)。免疫组织化学法检测肿瘤组织中树突细胞(DCs)的数量变化;采用酶联免疫吸附试验(ELISA)检测血清细胞因子的[干扰素(IFN)γ、白细胞介素(IL)4]水平;通过IFN-γ/IL-4比值估算Th1/Th2比值。组间比较采用单因素方差分析。 结果: 研究结果显示,冷冻消融联合GM-CSF对增加肿瘤内部DCs浸润数量具有协同作用,与其他组相比,冷冻+GM-CSF联合组治疗后第7天DCs数量最多[对照组(1.4±0.5)个/高倍视野,GM-CSF组(15.3±1.8)个/高倍视野,冷冻组(9.6±1.5)个/高倍视野,冷冻消融+GM-CSF组(17.4±1.3)个/高倍视野,F=138.60,P<0.05]。冷冻消融单独或联合GM-CSF可提高血清IFN-γ水平,降低血清IL-4水平。IFN-γ水平冷冻消融组从术前(45.6±2.4)pg/ml升高至(57.8±3.3)pg/ml,F=170.09,P<0.05,联合组从术前(45.6±2.4)pg/ml升高至(86.3±1.9)pg/ml,F=664.31,P<0.05;IL-4水平冷冻消融组从术前(7.21±0.63)pg/ml降低至(6.35±0.25)pg/ml,F=5.77,P<0.05,联合组从术前(7.21±0.63)pg/ml降低至(6.42±0.21)pg/ml,F=10.93,P<0.05。相反,GM-CSF降低血清IFN-γ水平,提高血清IL-4水平。冷冻消融和GM-CSF联合应用可显著提高Th1/Th2比值,冷冻消融+GM-CSF组从6.32±0.45升高至13.43±0.64,F=379.95,P<0.05。 结论: 冷冻消融联合GM-CSF治疗结直肠癌肝转移具有较好的抗肿瘤免疫反应,可增加肿瘤微环境中DCs的数量,提高抗肿瘤免疫应答。.

Publication types

  • English Abstract

MeSH terms

  • Animals
  • Colorectal Neoplasms* / immunology
  • Colorectal Neoplasms* / pathology
  • Colorectal Neoplasms* / therapy
  • Combined Modality Therapy
  • Cryosurgery*
  • Dendritic Cells / immunology
  • Female
  • Granulocyte-Macrophage Colony-Stimulating Factor* / therapeutic use
  • Interferon-gamma / blood
  • Interleukin-4 / blood
  • Liver Neoplasms* / immunology
  • Liver Neoplasms* / secondary
  • Liver Neoplasms* / therapy
  • Male
  • Mice
  • Mice, Inbred BALB C

Substances

  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Interleukin-4
  • Interferon-gamma