目的 探讨益气解毒方通过 miR-3127-5p 负调控信号转导及转录活化因子 2（STAT2）抑制鼻咽癌细胞增 殖、迁移和侵袭的分子机制。方法 动物实验：构建鼻咽癌裸鼠移植瘤模型，分为溶剂对照组、益气解毒方 组、5-氟尿嘧啶组，每组 5 只。通过苏木素-伊红染色观察鼻咽癌组织病理形态；免疫组织化学法和 Simple Western 法检测鼻咽癌组织中增殖和转移相关蛋白的表达水平。细胞实验一：将过表达 miR-3127-5p 的鼻咽癌 细胞，分为 oe-miR-NC 组、oe-miR-NC+益气解毒方组、oe-miR-3127-5p 组、oe-miR-3127-5p+益气解毒方 组，用益气解毒方（0.5 mg·mL-1 ）进行干预。实时无标记细胞功能分析仪（RTCA）监测细胞增殖。创伤愈合实验 及 Matrigel 侵袭小室法检测细胞迁移和侵袭能力的变化。Western Blot 法检测蛋白表达水平。细胞实验二：在 同时过表达 miR-3127-5p 和 STAT2 的基础上采用益气解毒方（0.5 mg·mL-1 ）干预，分为 oe-miR-NC+oe-NC 组、oe-miR-NC+oe-NC+益气解毒方组、oe-miR-3127-5p+oe-NC 组、oe-miR-3127-5p+oe-NC+益气解毒方 组、oe-miR-3127-5p+oe-STAT2 组、oe-miR-3127-5p+ oe-STAT2+益气解毒方组。RTCA 监测细胞增殖。创伤 愈合实验及 Matrigel 侵袭小室法检测细胞迁移和侵袭能力的变化。结果 苏木素-伊红染色发现益气解毒方组 鼻咽癌组织结构紊乱，甚至出现坏死、空泡等现象，同时益气解毒方降低了鼻咽癌组织中 XIAP、PCNA、 survivin、ID1、occludin、vimentin、claudin1、F-actin 的表达水平（P＜0.05，P＜0.01）。与 oe-miR-NC 组比较， 益气解毒方降低了鼻咽癌细胞的相对增殖率、迁移率和侵袭率（P＜0.05，P＜0.01），并降低了鼻咽癌细胞中 XIAP、PCNA、survivin、vimentin、F-actin、ANXA1 的表达水平（P＜0.05，P＜0.01）。与 oe-miR-NC+益气解 毒方组比较，miR-3127-5p+益气解毒方组的细胞相对增殖率、迁移率和侵袭率明显降低（P＜0.05，P＜0.01）， 并且鼻咽癌细胞中 XIAP、PCNA、survivin、vimentin、F-actin、ANXA1 的表达水平均下降（P＜0.05，P＜ 0.01）。通过析因设计分析发现益气解毒方和 miR-3127-5p 在抑制鼻咽癌细胞增殖、迁移和侵袭过程中存在一 定的交互作用。在过表达 miR-3127-5p 和 STAT2 的基础上采用益气解毒方进行干预，与 oe-miR-3127-5p+oeNC+益气解毒方组比，在 S18 细胞中，oe-miR-3127-5p+oe-STAT2+益气解毒方组的相对增殖率在 24 h 的上升 具有统计学意义（P＜0.05），在 36 、48 h 的上升没有统计学意义（P＞0.05）；5-8F 细胞中，oe-miR-3127-5p+ oe-STAT2+益气解毒方组的相对增殖率在 24 、36 、48 h 的上升均没有统计学意义（P＞0.05）；细胞相对迁移 率和侵袭率均有明显增加（P＜0.01）。结论 益气解毒方能够通过 miR-3127-5p 负调控 STAT2，最终抑制鼻咽 癌细胞增殖、迁移和侵袭。

Objective To investigate the molecular mechanism by which Yiqi Jiedu Formula suppresses proliferation， migration， and invasion of nasopharyngeal carcinoma cells via miR-3127-5p-mediated negative regulation of signal transducer and activator of transcription 2（STAT2）. Methods Animal experiments： A nasopharyngeal carcinoma xenograft model was established in nude mice and divided into solvent control group，Yiqi Jiedu Formula group，and 5- fluorouracil group，with 5 mice per group. Hematoxylin-eosin staining was used to observe histopathological morphology of nasopharyngeal carcinoma tissues. Immunohistochemistry and Simple Western assays were employed to assess the expression levels of proliferation- and metastasis-related proteins in tumor tissues. Cell experiment 1：Nasopharyngeal carcinoma cells overexpressing miR-3127-5p were treated with Yiqi Jiedu Formula （0.5 mg·mL-1 ） and divided into oemiR-NC group， oe-miR-NC + Yiqi Jiedu Formula group， oe-miR-3127-5p group， and oe-miR-3127-5p + Yiqi Jiedu Formula group. Real-time cellular analysis （RTCA） was performed to evaluate cell proliferation. Wound healing assay and Matrigel invasion chamber assay were used to assess changes in cell migration and invasion abilities. Western Blot was conducted to determine protein expression levels. Cell experiment 2：Cells overexpressing both miR-3127-5p and STAT2 were treated with Yiqi Jiedu Formula （0.5 mg·mL-1 ） and divided into oe-miR-NC + oe-NC group，oemiR-NC + oe-NC + Yiqi Jiedu Formula group，oe-miR-3127-5p + oe-NC group，oe-miR-3127-5p + oe-NC + Yiqi Jiedu Formula group，oe-miR-3127-5p + oe-STAT2 group，and oe-miR-3127-5p + oe-STAT2 + Yiqi Jiedu Formula group. RTCA was used to measure cell proliferation，while wound healing and Matrigel invasion assays were employed to evaluate migration and invasion capacities. Results Hematoxylin-eosin staining revealed that the Yiqi Jiedu Formula group exhibited disorganized nasopharyngeal carcinoma tissue structure，accompanied by necrosis and vacuolation. Yiqi Jiedu Formula also reduced the expression levels of XIAP，PCNA，survivin，ID1，occludin，vimentin，claudin1， and F-actin in nasopharyngeal carcinoma tissues （P＜0.05，P＜0.01）. Compared with the oe-miR-NC group，Yiqi Jiedu Formula decreased the relative proliferation rate，migration rate，and invasion rate of nasopharyngeal carcinoma cells （P＜0.05，P＜0.01），and reduced the expression levels of XIAP，PCNA，survivin，vimentin，F-actin，and ANXA1（P＜0.05，P＜0.01）. Compared with the oe-miR-NC + Yiqi Jiedu Formula group，the miR-3127-5p + Yiqi Jiedu Formula group showed significantly lower relative proliferation，migration，and invasion rates （P＜0.05，P＜ 0.01），along with decreased expression of XIAP，PCNA，survivin，vimentin，F-actin，and ANXA1 in nasopharyngeal carcinoma cells （P＜0.05， P＜0.01）. Factorial design analysis indicated an interactive effect between Yiqi Jiedu Formula and miR-3127-5p in inhibiting proliferation， migration， and invasion of nasopharyngeal carcinoma cells. Intervention with Yiqi Jiedu Formula in cells overexpressing both miR-3127-5p and STAT2 demonstrated that， compared with the oe-miR-3127-5p + oe-NC + Yiqi Jiedu Formula group，the oe-miR-3127-5p + oe-STAT2 + Yiqi Jiedu Formula group exhibited a statistically significant increase in relative proliferation rate at 24 h in S18 cells （P＜ 0.05）， while no statistical significance was observed at 36 h and 48 h （P＞0.05）. In 5-8F cells， the increase in relative proliferation rate at 24 h，36 h，and 48 h was not statistically significant （P＞0.05），but the relative migration and invasion rates were significantly elevated （P＜0.01）. Conclusion Yiqi Jiedu Formula inhibits proliferation， migration， and invasion of nasopharyngeal carcinoma cells through miR-3127-5p-mediated negative regulation of STAT2.

R285.5

湖南省自然科学基金项目（2023JJ30449，2024JJ6343）；湖南省教育厅资助项目（23A0298，23B0361）；湖南省卫生健康高层次人才 重大专项（R2023111）；湖南省卫健委项目（NO.D202307017740）；长沙市科技局指导项目（kzd22008）；湖南中医药大学校级课题（Z2023XJYB08）。