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目的 探讨呼肠孤病毒(Reo)是否通过CXCL10/CXCR3轴调控自然杀伤细胞(NK细胞)活化及向肿瘤细胞迁移。方法 将体外扩增的人源NK细胞分为Reo感染组(NK+Reo)和未感染对照组(NK),采用转录组测序技术(RNA-seq)检测两组细胞的差异表达基因，分析与NK细胞迁移相关的趋化因子受体CXCR3的表达变化；采用流式细胞术分析NK+Reo组中CXCR3和CD69的表达水平，免疫荧光染色检测NK细胞的CXCR3表达水平；采用定量聚合酶链反应(qPCR)和蛋白质印迹技术检测NK+Reo组CXCR3在mRNA和蛋白水平的表达量，qPCR和酶联免疫吸附试验(ELISA)检测Reo感染的结直肠癌细胞(Reo-DLD-1)中CXCL10的mRNA表达和CXCL10分泌情况；采用Transwell迁移实验评估阻断CXCR3后NK+Reo细胞向肿瘤细胞迁移和侵袭能力的变化。结果 与NK组相比，NK+Reo组存在多个差异表达基因，其中趋化因子受体CXCR3的上调最为显著(P<0.05);与DLD-1细胞相比，Reo感染后的Reo-DLD-1细胞CXCL10分泌水平显著升高(P<0.05);阻断CXCR3或CXCL10可抑制NK+Reo细胞向肿瘤细胞的迁移和侵袭能力。结论 Reo可以通过上调CXCL10/CXCR3轴的信号传导调控NK细胞向肿瘤细胞的迁移，从而增强NK细胞的抗肿瘤活性。
Abstract：Objective To investigate whether reovirus(Reo) regulates natural killer(NK) cell activation and migration toward tumor cells via the CXCL10/CXCR3 axis. Methods Human NK cells expanded in vitro were divided into a reovirus-infected group(NK+Reo) and an uninfected control group(NK). RNA sequencing(RNA-seq) was performed to identify differentially expressed genes between the two groups, with a focus on the chemokine receptor CXCR3 associated with NK cell migration. The expression levels of CXCR3 and CD69 in the NK+Reo group were analyzed by flow cytometry, and CXCR3 expression in NK cells was detected by immunofluorescence staining. Quantitative polymerase chain reaction(qPCR) and Western blotting were used to measure CXCR3 expression at both mRNA and protein levels in the NK+Reo group. The mRNA expression and secretion of CXCL10 in reovirus-infected colorectal cancer cells(Reo-DLD-1) were detected by qPCR and enzyme-linked immunosorbent assay(ELISA). Transwell migration assays were performed to evaluate changes in the migration and invasion capabilities of NK+Reo cells toward tumor cells following CXCR3 blockade. Results Compared with the NK group, the NK+Reo group exhibited multiple differentially expressed genes, among which the chemokine receptor CXCR3 was the most significantly upregulated(P<0.05). Compared with DLD-1 cells, Reo-DLD-1 cells showed significantly increased CXCL10 secretion after reovirus infection(P<0.05). Blockade of CXCR3 or CXCL10 inhibited the migration and invasion of NK+Reo cells toward tumor cells. Conclusion Reovirus can regulate NK cell migration toward tumor cells by upregulating signaling through the CXCL10/CXCR3 axis, thereby enhancing the anti-tumor activity of NK cells.
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基本信息:
DOI：10.19367/j.cnki.2096-8388.2026.05.001
中图分类号:R730.51
引用信息:
[1]袁代莎,杨再玲,龙世琪,等.呼肠孤病毒通过CXCL10/CXCR3轴促进NK细胞活化及迁移的实验研究[J].贵州医科大学学报,2026,51(05):625-634+644.DOI:10.19367/j.cnki.2096-8388.2026.05.001.
基金信息:
国家自然科学基金项目(82060564)
2026-05-20
2026-05-20