阿霉素抗4T1乳腺癌荷瘤小鼠的免疫调控机制

Immunomodulatory effect of adriamycin in mouse 4T1 model of breast cancer

摘要:

为了探讨阿霉素 (Adriamycin，ADM) 对4T1乳腺癌荷瘤鼠BALB/c的免疫调节作用，采用定量蛋白质组学串联质量标签 (TMT) 标记技术检测ADM对4T1乳腺癌差异蛋白的影响，利用多重数据库对差异蛋白进行生物信息学分析。根据蛋白质组学结果，寻找差异蛋白中与免疫调节功能相关的靶点，通过酶联免疫吸附测定 (Enzyme linked immunosorbent assay，ELISA) 观察ADM对乳腺癌组织中Th1细胞 (Helper T cells 1，Th1) 和Th2细胞 (Helper T cells 2，Th2) 的影响；通过流式细胞术分析ADM对CD4+ T细胞、CD8+ T细胞和调节性T细胞 (Regulatory T cells，Tregs) 的影响；HE染色观察ADM对4T1乳腺癌荷瘤鼠胸腺的改变。ADM上调170种差异蛋白，下调58种差异蛋白。有73种差异蛋白与免疫调控过程相关，KEGG (Kyoto encyclopedia of genes and genomes，KEGG) 富集于细胞因子及受体相关的重要蛋白通路、白介素17 (Interleukin 17，IL-17) 通路和癌症的转录调控通路。与免疫功能相关的差异蛋白与CD4+ T细胞、CD8+ T细胞和Tregs细胞的功能有关，而这些细胞的分型影响乳腺癌的预后。ADM极显著升高白介素2 (Interleukin 2，IL-2)，CD4+ T细胞、CD8+ T淋巴细胞含量 (P<0.01)，显著降低Tregs细胞含量 (P<0.05)。ADM抗乳腺癌的免疫调节蛋白有Ighm、Igkc、S100A8、S100A9和Tmsb4x。

Abstract:

To explore the immunomodulatory effect of adriamycin on 4T1 breast cancer. We used a tandem mass tag-based quantitative proteomic method to detect differential proteins in breast cancer tissues, and multiple bioinformatics databases to analyze the differentially expressed proteins in the proteome. Also, we used enzyme-linked immunosorbent assay to detect the effects of adriamycin on helper T cells 1 and 2 in breast cancer tissues, and flow cytometry to detect CD4+ T cells, CD8+ T cells and regulatory T cells. We discovered the immunomodulatory targets of adriamycin in differential proteins. In total 170 differential proteins were significantly up-regulated, whereas 58 were markedly down-regulated. In addition, 73 proteins were involved in immune regulation. Kyoto encyclopedia of genes and genomes enriched important protein pathways related to cytokines and factor receptors, interleukin 17 pathway and cancer transcriptional regulatory pathways. These pathways and important differential proteins related to immunomodulatory functions were ultimately regulated by adriamycin on CD4+ T cells, CD8+ T cells and regulatory T cells, thereby affecting the prognosis of breast cancer. Moreover, adriamycin significantly increased interleukin 2, CD4+ T and CD8+ T (P<0.01) and markedly reduced regulatory T cells (P<0.05). The function of adriamycin against triple-negative breast cancer was closely related to the immunoregulation process of the differential proteins Ighm, Igkc, S100A8, S100A9 and Tmsb4x. Adriamycin could regulate the content of helper T cells 1 cytokines, CD4+ T and CD8+ T lymphocytes in breast cancer and reduce the number of regulatory T cells to produce immunomodulatory effects.