脾多肽对乳腺癌荷瘤小鼠的抑瘤效应及肿瘤免疫微环境影响的实验研究  

Experimental study on the inhibitory effect of spleen polypeptide on tumor-bearing mice with breast cancer and the influence of tumor immune microenvironment

作　　者：章钰辰 严雪绮 蒋梦萍 付子毅[2] 殷咏梅[2] ZHANG Yuchen;YAN Xueqi;JIANG Mengping;FU Ziyi;YIN Yongmei(Department of Oncology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China)

机构地区：[1]南京医科大学第一附属医院肿瘤科,南京210029 [2]南京医科大学附属第一医院妇幼研究中心/乳腺病研究中心,210029

出　　处：《临床肿瘤学杂志》2022年第4期297-302,共6页Chinese Clinical Oncology

基　　金：吴阶平基金资助项目(320.6750.17006)。

摘　　要：目的探讨脾多肽注射液对乳腺癌荷瘤小鼠抗肿瘤治疗的效果和安全性以及对乳腺癌肿瘤微环境中淋巴细胞亚群比例的影响。方法动物实验使用20只BALB/C小鼠进行皮下瘤造模。造模成功小鼠随机分为3组:对照组、脾多肽低剂量组和脾多肽高剂量组。脾多肽低剂量组和脾多肽高剂量组连续10天分别每天尾静脉注射1 mg/kg和10 mg/kg的脾多肽注射液,对照组小鼠经尾静脉注射等体积生理盐水。实验过程中记录小鼠体质量和肿瘤体积变化,实验结束后测量小鼠肿瘤组织的体积和质量并使用流式细胞术分析小鼠肿瘤组织单细胞悬液中不同类型淋巴细胞的比例。体外实验则建立人外周血单核细胞(PBMCs)和乳腺癌MDA-MB-231细胞共培养模型,实验组培养基中分别加入0.8μg/ml(脾多肽低剂量组)、8μg/ml(脾多肽高剂量组)的脾多肽注射液,对照组加入等量的完全培养基。处理48 h后收集共培养体系中的PBMCs,用流式细胞术分析PBMCs中淋巴细胞亚群比例变化。结果各组荷瘤小鼠的体质量和肿瘤体积均随着时间的推移增长。与对照组相比,不同剂量脾多肽治疗组的荷瘤小鼠在使用脾多肽治疗10天后,小鼠体质量变化率、肿瘤体积和肿瘤质量的差异无统计学意义(P>0.05)。与对照组相比,脾多肽治疗组能够上调乳腺癌荷瘤小鼠肿瘤组织中CD4^(+)T淋巴细胞和CD8^(+)T淋巴细胞的比例(P<0.05)。此外,对于MDA-MB-231细胞与PBMCs的共培养体系,低剂量和高剂量脾多肽均能够不同程度地上调PBMCs亚群中CD4^(+)T淋巴细胞和CD8^(+)T淋巴细胞的比例(P<0.05)。结论脾多肽能增强乳腺癌荷瘤小鼠的抗肿瘤免疫能力且对于小鼠无明显毒副作用并增加乳腺癌肿瘤免疫微环境中CD4^(+)T淋巴细胞和CD8^(+)T淋巴细胞的比例,具有进一步应用于临床研究中的价值。Objective To investigate the effect and safety of spleen polypeptide injection on the antitumor treatment of breast cancer bearing mice and the effect of spleen peptide injection on the lymphocyte subsets in the microenvironment of breast cancer.Methods Twenty BALB/C mice were used to induce subcutaneous tumor in animal experiments.The mice were randomly divided into three groups:normal control group,spleen polypeptide low-dose group and spleen polypeptide high-dose group.Mice in the spleen polypeptide low-dose group and high-dose group received 1 mg/kg or 10 mg/kg of spleen polypeptide injection per day via tail vein for 10 day.The control group mice were injected with the same volume of normal saline through the tail vein.Changes of body mass and tumor volume of mice were recorded during the experiment.The volume and mass of tumor tissue were measured at the end of the experiment,and the proportions of different types of lymphocytes in single cell suspension of mouse tumor tissue were analyzed by flow cytometry.The co-culture model of human peripheral blood monocytes(PBMCs)and MDA-MB-231 cells was established in vitro.Additionally,0.8μg/ml(low-dose group)and 8μg/ml(high-dose group)spleen polypeptide injection were added to the culture medium of the experimental group,and the same amount of complete medium was added to the control group.After 48 hours of treatment,PBMCs in co-culture system were collected and the proportions of lymphocyte subsets in PBMCs were analyzed by flow cytometry.Results The body mass and tumor volume of tumor-bearing mice in each group increased with time.Compared with the control group,there was no significant difference in the change rate of body mass,tumor volume and tumor mass of tumor-bearing mice in different doses of spleen polypeptide treatment groups after 10 days of spleen polypeptide treatment(P>0.05).Compared with the control group,the spleen polypeptide treatment could upregulate the proportions of CD4^(+)T lymphocytes and CD8^(+)T lymphocytes in tumor tissues of tumor-bea

关 键 词：乳腺癌 脾多肽 肿瘤免疫微环境 外周血单核细胞 

分 类 号：R737.9[医药卫生—肿瘤]