冷冻消融联合GM-CSF治疗结直肠癌肝转移的试验研究
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摘要
背景：冷冻消融技术逐渐发展成为结直肠癌肝转移重要的治疗手段,冷冻免疫反应研究将冷冻消融局部治疗与全身免疫治疗有机结合起来,为肿瘤综合治疗提供理论和实验依据。目的：评价冷冻消融治疗结直肠癌肝转移后对机体抗肿瘤免疫状态的影响,探讨冷冻消融联合GM-CSF治疗结直肠癌肝转移的可行性与有效性。材料与方法：复制小鼠结肠癌CT-26细胞肝转移模型,将动物随机分为对照组、GM-CSF治疗组、冷冻治疗组、冷冻+GM-CSF治疗组。利用Endocare公司氩氦冷冻系统直径1.7mm冷冻针行冷冻消融治疗。观察肝转移瘤大小及荷瘤小鼠生存期,免疫组织化学评价肿瘤局部DC的数量；ELISA法检测外周血IFN-γ、、IL-4水平,以IFN-γ/IL-4计算Th1/Th2比值。结果：(1)冷冻消融+GM-CSF治疗可明显缩小结直肠癌肝转移病灶大小(P<0.001),并延长荷瘤小鼠生存期(p<0.001)。(2)冷冻+GM-CSF治疗术前、术后7天、术后14天、术后21天肿瘤局部DC数量分别为每高倍视野下(1.5±0.55)个、(17.4±1.26)、(14.6±0.54)、(7.2±1.47)个(P<0.05)。(3)冷冻+GM-CSF治疗术前、术后7天、术后14天、术后21天外周血Th1/Th2比值分别为6.32±0.45、13.43±0.23、14.87±0.53、15.54±0.64(P<0.05)。结论：冷冻消融联合GM-CSF治疗可减小结直肠癌肝转移灶大小,并延长荷瘤小鼠生存期,冷冻联合GM-CSF治疗可增加肿瘤组织局部DC浸润,提高Th1/Th2比值。
Backgroud:Cryoablation is becoming the primary therapy for liver metastasis from colorectal cancer, while the exploration of cryoimmunology response will provide the combination of local therapy with immune therapy, and would provide rationale and experimental foundation for synthetic therapy of advanced cancer. Objective:To make the models of liver metastasis from colorectal cancer. To assess the anti-tumor immune response after cryoablation combined GM-CSF treatment. Material and
Methods:Mouse models of liver metastasis from colorectal cancer were made, and the tumor-bearing mice were divided into different groups:control group, GM-CSF group, cryoablation group, cryoablation+GM-CSF group. Cryoablation was applied by cryosurgery needle with1.7mm diameter from Endocare Argon-Helium cryosurgery system. Tumor size and survival time were observed, the number changes of dendritic cells in tumor were assessed by immunohistochemistry, serum cytokine (IFN-y, IL-4) levels were analyzed by enzyme-linked immunosorbent assay (ELISA), and the Thl/Th2ratio was estimated from the IFN-y/IL-4ratio.
Result:(1) The Cryoablation+GM-CSF treatment significantly reduced to tumor lesion size (P＜0.001), and to extend the survival time of tumor-bearing mice (p＜0.001).(2) On the day before, seventh day after,14th day after,21st day after cryoablation+GM-CSF treatment, the DC numbers per high power field in tumor were1.5±0.55,17.4±1.26,14.6±0.54,7.2±1.47respectively (P＜0.05).(3) The Thl/Th2ratios were6.32±0.45,13.43±0.23,14.87±0.53,15.54±0.64on the day before, seventh days after,14th day after,21st day after cryoablation+GM-CSF treatment (P＜0.05). Conclusion:Combined cryoablation with GM-CSF treatment could increase the number of DC in tumor micro environment, improve the systemic anti-tumor immune response, increase the ratio of Thl/Th2, prolong the median survival times.
Backgroud:Cryoablation is becoming the primary therapy for liver metastasis from colorectal cancer, while the exploration of cryoimmunology response will provide the combination of local therapy with immune therapy, and would provide rationale and experimental foundation for synthetic therapy of advanced cancer. Objective:To make the models of liver metastasis from colorectal cancer. To assess the anti-tumor immune response after cryoablation combined GM-CSF treatment. Material and
Methods:Mouse models of liver metastasis from colorectal cancer were made, and the tumor-bearing mice were divided into different groups:control group, GM-CSF group, cryoablation group, cryoablation+GM-CSF group. Cryoablation was applied by cryosurgery needle with1.7mm diameter from Endocare Argon-Helium cryosurgery system. Tumor size and survival time were observed, the number changes of dendritic cells in tumor were assessed by immunohistochemistry, serum cytokine (IFN-y, IL-4) levels were analyzed by enzyme-linked immunosorbent assay (ELISA), and the Thl/Th2ratio was estimated from the IFN-y/IL-4ratio.
Result:(1) The Cryoablation+GM-CSF treatment significantly reduced to tumor lesion size (P＜0.001), and to extend the survival time of tumor-bearing mice (p＜0.001).(2) On the day before, seventh day after,14th day after,21st day after cryoablation+GM-CSF treatment, the DC numbers per high power field in tumor were1.5±0.55,17.4±1.26,14.6±0.54,7.2±1.47respectively (P＜0.05).(3) The Thl/Th2ratios were6.32±0.45,13.43±0.23,14.87±0.53,15.54±0.64on the day before, seventh days after,14th day after,21st day after cryoablation+GM-CSF treatment (P＜0.05). Conclusion:Combined cryoablation with GM-CSF treatment could increase the number of DC in tumor micro environment, improve the systemic anti-tumor immune response, increase the ratio of Thl/Th2, prolong the median survival times.
引文
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