中国医药导报
中国医药期刊欢迎您 今天是   2024年12月30日星期一
设为首页 | 加入收藏 
 
        首 页      期刊介绍      下载中心      关于本刊      投稿指南      期刊订阅      在线留言      广告合作      联系我们      返回中国当代医药网
中国医药导报  2023, Vol. 20 Issue (33): 28-33    DOI: 10.20047/j.issn1673-7210.2023.33.05
  论著 本期目录 | 过刊浏览 | 高级检索 |
放疗、吉西他滨联合免疫检查点抑制剂的三重疗法对三阴性乳腺癌肿瘤微环境的作用机制
吴苗苗1  房坤2
1.中国科学技术大学附属第一医院西区 安徽省肿瘤医院 安徽省立医院西区肿瘤表观遗传学研究室,安徽合肥 230031;
2.中国科学技术大学附属第一医院 安徽省立医院生物医学工程处,安徽合肥 230001
Mechanism of action of triple therapy with radiotherapy and Gemcitabine combined with immune checkpoint inhibitors on the microenvironment of triple-negative breast cancer
WU Miaomiao1 FANG Kun2
1.Tumor Epigenetics Lab, West Branch of the First Affiliated Hospital of University of Science and Technology of China Anhui Provincial Cancer Hospital West Branch of Anhui Provincial Hospital, Anhui Province, Hefei 230031, China;
2.Department of Biomedical Engineering, the First Affiliated Hospital of University of Science and Technology of China Anhui Provincial Hospital, Anhui Province, Hefei 230001, China
全文: PDF (1559 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 目的 探讨放疗、吉西他滨(Gem)联合免疫检查点抑制剂的三重疗法对三阴性乳腺癌肿瘤微环境的影响,并揭示其可能的发生机制。 方法 选取29只雌性5周龄体重18~20 g的SPF级BALB/c小鼠,实验第1天在其左腹部皮下注射约1×106个4T1细胞,建立三阴性乳腺癌移植瘤模型。待肿瘤体积增长至100 mm3时采用随机数字表法将其分为对照组(5只)、程序性死亡受体配体1抑制剂(aPD-L1)组(6只)、放疗(RT)+aPD-L1组(6只)、Gem+aPD-L1组(6只)和三联组(6只)。除对照组外,其余四组在实验第6、8、10天于瘤内注射药物(Gem 5 mg/kg,aPD-L1 3 mg/kg),每次注射药物24 h后,对RT+aPD-L1组、三联组进行RT(2 Gy)。实验第30天处死动物,称量各组瘤重,并对主要器官进行苏木精-伊红染色。采用流式细胞术检测各组外周血及肿瘤中浸润的CD3+CD8+T细胞绝对百分比。酶联免疫吸附试验检测各组血清和瘤内γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的表达。免疫组织化学染色检测各组瘤中Ki-67、微血管密度(MVD)-CD31和γH2AX的表达,TUNEL检测评价细胞凋亡情况。Western blot检测各组瘤中血管内皮生长因子(VEGF)和低氧诱导因子-1α(HIF-1α)的表达。 结果 实验过程中各组体重比较,差异无统计学意义(P>0.05)。实验结束时,RT+aPD-L1组、三联组瘤重均低于aPD-L1组,三联组低于RT+aPD-L1组(P<0.05)。苏木精-伊红染色染色显示各组心脏、肝脏、脾脏、肾脏没有明显损伤。aPD-L1组外周血CD3+CD8+ T细胞绝对百分比、TNF-α、IL-6高于对照组;RT+aPD-L1组外周血IL-6高于aPD-L1组,三联组外周血CD3+CD8+ T细胞绝对百分比、IFN-γ、IL-6高于aPD-L1组;三联组外周血CD3+CD8+ T细胞绝对百分比高于RT+aPD-L1组、Gem+aPD-L1组,IL-6高于Gem+aPD-L1组(P<0.05)。aPD-L1组肿瘤组织中CD3+CD8+ T细胞绝对百分比、IFN-γ、TNF-α、IL-6高于对照组;RT+aPD-L1组、Gem+aPD-L1组、三联组肿瘤组织中CD3+CD8+ T细胞绝对百分比高于aPD-L1组,三联组TNF-α、IL-6高于aPD-L1组;三联组肿瘤组织中CD3+CD8+ T细胞绝对百分比、IL-6高于RT+aPD-L1组、Gem+aPD-L1组,IFN-γ高于RT+aPD-L1组(P<0.05)。免疫组织化学染色和TUNEL检测显示三联组肿瘤组织中Ki-67的表达减少,凋亡的细胞增多。三联组MVD-CD31低于aPD-L1组,γH2AX高于aPD-L1组、Gem+aPD-L1组(P<0.05)。aPD-L1组VEGF低于对照组,RT+aPD-L1组、Gem+aPD-L1组、三联组VEGF低于aPD-L1组,三联组HIF1α低于aPD-L1组和RT+aPD-L1组(P<0.05)。 结论 在小鼠三阴性乳腺癌移植瘤模型中应用放疗、Gem联合免疫检查点抑制剂的三重疗法可以延缓肿瘤生长,提高外周血和肿瘤中CD3+CD8+T细胞绝对百分比,促进血清和肿瘤中炎症因子的释放,还可以改善瘤内缺氧状况,抑制肿瘤微血管生长。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
吴苗苗1  房坤2
关键词 三阴性乳腺癌三重疗法免疫应答肿瘤微环境    
Abstract:Objective To investigate the effect of triple therapy of radiotherapy, Gemcitabine (Gem) combined with immune checkpoint inhibitors on the microenvironment of triple-negative breast cancer, and to reveal its possible mechanism. Methods A total of 29 SPF BALB/c mice aged five weeks and weighing 18-20 g were selected. On the first day of the experiment, about 1×106 4T1 cells were injected subcutaneously into the left abdomen to establish triple-negative breast cancer transplant tumor model. When the tumor volume increased to 100 mm3, the mice were divided into control group (five mice), anti-programmed death receptor ligand 1 (aPD-L1) group (six mice), radiotherapy (RT) +aPD-L1 group (six mice), Gem +aPD-L1 group (six mice), and triple group (six mice) by random number method. In addition to the control group, the other four groups were injected with drugs (Gem 5 mg/kg, aPD-L1 3 mg/kg) in the tumor at the 6th, 8th, and 10th days of the experiment. Twenty-four hours after each injection, RT (2 Gy) was administered to the RT+aPD-L1 group and the triple group. On the 30th day of the experiment, the animals were killed, the tumor weight of each group was weighed, and the main organs were stained with hematoxylin-eosin. The absolute percentage of CD3+CD8+ T cells in peripheral blood and tumor was detected by flow cytometry. The expression of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) in peripheral blood and tumor were detected by enzyme-linked immunosorbent assay. The expressions of Ki-67, microvessel density(MVD)-CD31, and γH2AX were detected by immunohistochemistry, and apoptosis was evaluated by TUNEL detection. The expression of vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1 α (HIF-1α) were detected by Western blot analysis. Results There was no significant difference in body weight among all groups (P>0.05). At the end of the experiment, the tumor weight of RT+aPD-L1 group and triple group were lower than those of aPD-L1 group, and those of triple group were lower than those of RT+aPD-L1 group (P<0.05). Hematoxylin-eosin staining showed no obvious damage to the heart, liver, spleen, and kidney in each group. The absolute percentage of CD3+CD8+ T cells , TNF-α, and IL-6 in peripheral blood of aPD-L1 group were higher than those of control group. Peripheral blood IL-6 in RT+aPD-L1 group was higher than that in aPD-L1 group, and absolute percentage of CD3+CD8+ T cells , IFN-γ, and IL-6 of peripheral blood in triple group were higher than those in aPD-L1 group. The absolute percentage of CD3+CD8+ T cells of peripheral blood in triple group was higher than that of RT+aPD-L1 group and Gem+aPD-L1 group, and IL-6 was higher than that of Gem+aPD-L1 group (P<0.05). The absolute percentage of CD3+CD8+ T cells , IFN-γ, TNF-α, and IL-6 of tumor in aPD-L1 group were higher than those in control group. The absolute percentages of CD3+CD8+ T cells of tumor in RT+aPD-L1 group, Gem+aPD-L1 group, and triple group were higher than those in aPD-L1 group, and TNF-α and IL-6 of tumor in triple group were higher than those in aPD-L1 group. The absolute percentage of CD3+CD8+ T cells and IL-6 of tumor in triple group were higher than those in RT+aPD-L1 group and Gem+aPD-L1 group, and IFN-γ was higher than those in RT+aPD-L1 group (P<0.05). Immunohistochemical staining and TUNEL detection showed that the expression of Ki-67 decreased and the apoptotic cells increased in the tumor tissues of triple group. MVD-CD31 in triple group was lower than that in aPD-L1 group, γH2AX was higher than that in aPD-L1 group and Gem+aPD-L1 group (P<0.05). VEGF in aPD-L1 group was lower than that in control group, VEGF in RT+aPD-L1 group, Gem+aPD-L1 group, and triple group was lower than that in aPD-L1 group, and HIF-1α in triple group was lower than that in aPD-L1 group and RT+aPD-L1 group (P<0.05). Conclusion The triple therapy of radiotherapy, Gem combined with immune checkpoint inhibitors can delay tumor growth, increase the absolute percentage of CD3+CD8+ T cells in peripheral blood and tumor, promote the release of inflammatory factors in serum and tumor, improve intratumor hypoxia and inhibit tumor microvascular growth in mouse triple-negative breast cancer transplant tumor model.
Key wordsTriple-negative breast cancer    Triple therapy    Immune response    Tumor microenvironment
    
基金资助:安徽省自然科学基金项目(1808085MH296)。
通讯作者: 房坤(1979.7-),男,博士,高级工程师;研究方向:纳米制剂及其在生物医学领域的应用。   
作者简介: 吴苗苗(1989.1-),女,硕士;研究方向:纳米制剂在乳腺癌诊疗中的应用。
引用本文:   
吴苗苗1  房坤2. 放疗、吉西他滨联合免疫检查点抑制剂的三重疗法对三阴性乳腺癌肿瘤微环境的作用机制[J]. 中国医药导报, 2023, 20(33): 28-33.
WU Miaomiao1 FANG Kun2. Mechanism of action of triple therapy with radiotherapy and Gemcitabine combined with immune checkpoint inhibitors on the microenvironment of triple-negative breast cancer. 中国医药导报, 2023, 20(33): 28-33.
链接本文:  
https://www.yiyaodaobao.com.cn/CN/10.20047/j.issn1673-7210.2023.33.05     或     https://www.yiyaodaobao.com.cn/CN/Y2023/V20/I33/28

 

中华人民共和国互联网药品信息服务资格证书(京)-非经营性-2016-0092
京ICP11001767号-2  京公网安备 11010502046598号  期刊出版许可证 广告经营许可证
信息产业部备案管理系统网址 http://beian.miit.gov.cn/state/outPortal/loginPortal.action
投稿热线:010-59626203
本刊地址:北京市朝阳区东四环中路78号楼(大成国际中心B1座)8B02室 邮编:100124
传真:010-59626204 投稿信箱:ddyy@vip.163.com
版权所有:中文传媒集团北京期刊有限公司
技术支持:北京玛格泰克科技发展有限公司