目的：基于蛋白质组学技术探讨麻疹减毒活疫苗191 株（MV-Hu191）在体内外对三阴性乳腺癌MDA-MB-231、4T1细胞的影响及其作用机制。方法：采用CCK-8法检测MV-Hu191 对MDA-MB-231 和4T1细胞增殖的影响；液相色谱-质谱联用技术分析MV-Hu191 处理对MDA-MB-231 细胞中蛋白质谱的影响，多重数据库筛选蛋白质谱中的典型差异蛋白质并进行GO、 KEGG、亚细胞定位与功能注释。瘤内注射1×106 TCID50 MV-Hu191 干预4T1细胞移植瘤模型小鼠，流式细胞术检测小鼠脾组织中T细胞亚群，ELISA 法检测小鼠血清TNF-α和IL-6含量。结果：体外实验结果表明，MV-Hu191 具有抑制MDA-MB-231 和4T1细胞增殖的作用，差异均具有统计学意义（P<0.01）。蛋白质组学分析结果显示，MV-Hu191 作用MDA-MB-231 细胞后明显上调蛋白质有38个、下调有12个；差异表达的蛋白质主要参与细胞黏附、信号受体激活、细胞代谢、应激反应等生物学过程，22个差异蛋白质亚细胞定位位于细胞外，KEGG功能分类显示与免疫调节功能相关的差异蛋白质最多且均为上调蛋白，包括C4A、C8B、 SERPINF2、A2M、SERPINC1、CTSB、SERPING1、C5；PPI 预测发现免疫相关差异蛋白与CD4、CD8、TNF-α 及IL-6 相互关联。体内实验结果显示，MV-Hu191 干预组小鼠脾组织中CD4+ T 细胞数量略高于对照组，但差异无统计学意义（P>0.05），CD4+/CD8+ T细胞比值明显高于对照组（P<0.05），血清TNF-α和IL-6含量显著上升（均P<0.01）。结论：MV-Hu191 显著抑制MDA-MB-231、 4T1细胞增殖及拮抗4T1细胞荷瘤小鼠成瘤性，其机制可能是MV-Hu191通过激活免疫效应分子实现抗肿瘤作用。

Objective: To investigate the effect and mechanism of live attenuated measles vaccine strain 191 (MV-Hu191) on triple negative breast cancer MDA-MB-231 and 4T1 cells in vitro and in vivo based on proteomics. Methods: CCK-8 method was used to analyze the effect of MV-Hu191 on the proliferation of MDA-MB-231 and 4T1 cells. Liquid chromatography-mass spectrometry was used to analyze the effect of MV-Hu191 treatment on protein spectrum in MDA-MB-231 cells. Multiple databases were used to screen typical differentially expressed proteins, followed with GO, KEGG, subcellular localization and functional annotation. Intratumoral injection of 1×106 TCID50 MV-Hu191 was used to intervene the growth of 4T1 cell transplanted tumor in mice. Flow cytometry was applied to detect T cell subpopulations in splenic tissue, and ELISA was used for the detection of serum levels of TNF-α and IL-6. Results: In vitro experiments showed that MV-Hu191 could significantly inhibit the proliferation of MDA-MB-231 and 4T1 cells (P<0.01). Proteomic analysis showed that 38 proteins were significantly upregulated while 12 proteins were downregulated in MDA-MB-231 cells after MV-Hu191 treatment. The differentially expressed proteins were mainly involved in the biological processes of cell adhesion, signaling receptor activation, cell metabolism, and stress responses. The subcellular localization of 22 differentially expressed proteins was located outside the cell. The KEGG functional classification showed that the most differentially expressed proteins were related to immunomodulatory functions and they were all upregulated, including C4A, C8B, SERPINF2, A2M, SERPINC1, CTSB, SERPING1, and C5; PPI prediction found immune-related differential proteins were associated with CD4, CD8, TNF-ɑ and IL-6. In vivo experiments showed that the number of CD4+ T cells in spleen tissues of mice in the MV-Hu191 intervention group was higher than that in the control group, but the difference was not significant (P>0.05). The ratio of CD4+/CD8+ T cells and the serum levels of TNF-ɑ and IL-6 in the MV-Hu191 intervention group were significantly higher than those in the control group (P<0.05, P<0.01). Conclusion: MV-Hu191 significantly inhibits the proliferation of MDA-MB-231 and 4T1 cells and antagonizes the tumorigenicity in mice bearing 4T1 cell xenograft. The mechanism may be that MV-Hu191 achieve its anti-tumor effect by activating immune effector molecules.

省卫生健康委科学技术基金（No. GZWKJ2023-280）；2021 年国家级大学生创新创业训练项目（No. 202110660028）；2020 年省级大学生创新训练项目（No. S202010660053）；贵州省教育厅平台项目（No. 黔教技[2022]019 号）