首页> 中文期刊> 《南方医科大学学报》 >sPD-1过表达增强衰老肿瘤细胞疫苗抗小鼠乳腺癌作用

sPD-1过表达增强衰老肿瘤细胞疫苗抗小鼠乳腺癌作用

         

摘要

目的 探讨可溶性PD-1(sPD-1)过表达后对衰老肿瘤细胞疫苗(STCV)抗小鼠乳腺癌的增强作用.方法 干扰素γ(IFN-γ)刺激小鼠乳腺癌细胞4T1,流式细胞术检测PD-L1表达;sPD-1过表达慢病毒感染4T1,显微镜观察增强绿色荧光蛋白表达情况;CCK8实验比较4T1、4T1/sPD-1增殖活力;qRT-PCR、Western blot从mRNA、蛋白质水平验证sPD-1表达;干扰素γ预处理4T1细胞,添加4T1/sPD-1培养上清,孵育后流式细胞术检测PD-1阳性细胞比例;X射线照射联合Veliparib处理4T1、4T1/sPD-1细胞,衰老相关β-半乳糖苷酶(SA-β-gal)染色,观察蓝染细胞比例;Balb/c小鼠右后腿皮下种植4T1,左后腿注射PBS、4T1 STCV、4T1/sPD-1 STCV,观察各组小鼠成瘤率.结果 IFN-γ能导致4T1细胞PD-L1上调(P<0.001),且浓度越高,PD-L1表达上调越明显,最高达(84.80±1.03)%;病毒感染4T1细胞后,显微镜下可见绿色荧光;CCK8实验中4T1、4T1/sPD-1细胞增殖曲线无明显差异(P>0.05);4T1/sPD-1细胞在mRNA和蛋白质上均可检测到sPD-1表达产物,4T1细胞则均未能检测到;干扰素γ预处理的4T1细胞,PD-1阳性比例(6.893±0.271)%,添加4T1/sPD-1细胞培养液处理后,PD-1阳性细胞比例升高达(55.450±0.555)%(P<0.001);4T1、4T1/sPD-1在联合处理后,镜下见大量蓝染变形的衰老细胞;小鼠荷瘤实验中,预防肿瘤发生实验,PBS组所有小鼠出现肿瘤生长,4T1 STCV组有28.787%小鼠未发生肿瘤,4T1/sPD-1 STCV组近55.556%小鼠未发现肿瘤发生;治疗肿瘤实验中,观察期内PBS组小鼠均发生肿瘤,4T1 STCV、4T1/sPD-1 STCV组无瘤小鼠比例分别为11.111%和38.89%.结论 衰老肿瘤细胞疫苗对小鼠乳腺癌具有防治作用,且sPD-1过表达后能够增强衰老肿瘤细胞疫苗的抗肿瘤作用.%Objective To investigate whether soluble PD-1 overexpression in 4T1 senescence tumor cells enhances the anti-tumor effect of senescence tumor cell vaccine (STCV) against breast tumor cells in a tumor-bearing mouse model. Methods 4T1 cells were treated with interferon-γ (IFN-γ) and the expression of PD-L1 was detected by flow cytometry. CCK8 assay was used to compare the cell proliferation activity between 4T1 cells and 4T1 cells infected by a lentiviral vector of sPD-1 (4T1/sPD-1 cells), and the expressions of sPD-1 mRNA and protein in 4T1/sPD-1 cells were detected using qPCR and Western blotting. The culture supernatant of 4T1/sPD-1 cells was added in 4T1 cells pre-treated with IFN-γ, and PD-1-positive 4T1 cells were detected with flow cytometry. Senescenceβ-galactosidase staining kit was used to detect the senescent 4T1 and 4T1/sPD-1 cells following exposure to X-ray radiation and Veliparib. Balb/c mice bearing subcutaneous 4T1 tumor xenografts were treated with injections of PBS, 4T1 STCV, or 4T1/sPD-1 STCV, and tumor growth was observed. Results Stimulation with IFN-γconcentration-dependently up-regulated PD-L1 expression by as much as (84.80 ± 1.03)% in 4T1 cells (P<0.001). sPD-1 overexpression in 4T1 cells did not significantly affect the cell proliferation. Treatment of 4T1 cells with 4T1/sPD-1 cell culture supernatant significantly increased the proportion of PD-1 + cells from (6.893 ± 0.271)% to (55.450 ± 0.555)% (P<0.001). X-ray irradiation combined with Veliparib caused obvious senescence in 4T1 and 4T1/sPD-1 cells. In the tumor-preventing experiment, tumor formation occurred in all the mice in PBS group;28.787%of the mice in 4T1 STCV group and 55.556%in 4T1/sPD-1 STCV group showed no tumor formation. In the tumor treatment experiment, tumor formation occurred in all the mice in PBS groups while in 4T1 STCV and 4T1/sPD-1 STCV groups, 11.111% and 38.89% of the mice were tumor-free during the observation period, respectively. Conclusions Senescence tumor cells vaccine has anti-tumor effect against breast cancer in mice, and sPD-1 over-expression can enhance this effect of the vaccine.

著录项

  • 来源
    《南方医科大学学报》 |2018年第1期|20-26|共7页
  • 作者单位

    南方医科大学肿瘤中心//南方医科大学中西医结合医院,广东 广州 510315;

    南方医科大学肿瘤中心//南方医科大学中西医结合医院,广东 广州 510315;

    南方医科大学肿瘤中心//南方医科大学中西医结合医院,广东 广州 510315;

    南方医科大学普宁人民医院药剂科,广东 普宁 515300;

    南方医科大学肿瘤中心//南方医科大学中西医结合医院,广东 广州 510315;

    南方医科大学肿瘤中心//南方医科大学中西医结合医院,广东 广州 510315;

    南方医科大学南方医院肿瘤科,广东 广州 510515;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类
  • 关键词

    可溶性PD-1; X射线; Veliparib; 衰老肿瘤细胞疫苗; 乳腺癌;

相似文献

  • 中文文献
  • 外文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号