目的 基于生物信息学手段与实验验证探究补骨脂酚（Bak）对鼻咽癌细胞增殖、凋亡、迁移与侵袭的影 响与机制。方法 CCK-8 和实时细胞分析系统（RTCA）用于检测细胞增殖，PI 染色分析细胞周期，Annexin V-FITC/PI双荧光染色法、Hoechst 33342和JC-1线粒体膜染色检测细胞凋亡，创伤愈合实验和Transwell小室法 分别检测细胞迁移和侵袭能力。Western Blot法检测相关蛋白表达水平，网络药理学分析预测BAK潜在的目标 和假定途径之间的相互关系，分子对接检测核心靶标与BAK的结合亲和力。结果 CCK-8和RTCA结果显示， BAK可抑制鼻咽癌细胞增殖。PI染色结果显示，BAK干预鼻咽癌5-8F细胞36 h后，BAK（30 μmol·L-1）可使处 于 S期和 G2/M 期的 5-8F 细胞比例增加，处于 G1期的 6-10B 细胞比例增加，BAK（40 μmol·L-1）可使处于亚 G1 期和S期的5-8F细胞比例增加，处于G2/M期的6-10B细胞比例增加。Hoechst 33342、JC-1与Annexin V-FITC/ PI双荧光染色法结果显示，BAK可导致5-8F与6-10B细胞固缩、碎裂和核溶解，线粒体膜电位降低，细胞凋 亡率增加。创伤愈合实验和Transwell小室法结果则显示BAK可降低5-8F与6-10B迁移和侵袭率。Western Blot 结果显示，BAK 可降低 5-8F 与 6-10B 细胞增殖（PCNA）、细胞周期（Cyclin D3）、凋亡（XIAP）和迁移（Vimentin 和N-cadherin）蛋白的表达水平。网络药理学分析显示，鼻咽癌中MAPK信号通路受BAK影响最大，分子对接 的结果显示，BAK对核心靶基因产物VEGFA、PTGS2、EGFR和TNF具有亲和力，实验验证表明BAK可有效降 低5-8F与6-10B细胞中ERK1/2和p-ERK1/2的表达水平以及VEGFA、PTGS2、EGFR和TNF表达水平。结论 补 骨脂酚可抑制鼻咽癌细胞增殖，诱导凋亡，抑制细胞迁移和侵袭，其作用机制可能与影响 VEGFA、PTGS2、 EGFR和TNF等靶点以及MAPK信号通路有关。

Objective To explore the effect and mechanism of bakuchiol （BAK）on the proliferation， apoptosis， migration， and invasion of nasopharyngeal carcinoma cells based on bioinformatics methods and experimental verification. Methods CCK-8 and real-time cell analysis system（RTCA）were used to detect cell proliferation. PI staining was used to analyze cell cycle. Annexin V-FITC/PI dual fluorescence staining， Hoechst 33342 and JC-1 mitochondrial membrane staining were used to detect cell apoptosis， and wound healing experiments and Transwell chamber method were used to detect cell migration and invasion ability. Western Blot was used to detect the expression levels of relevant proteins，network pharmacology was used to predict the potential targets and hypothesized pathways of BAK，and molecular docking was used to detect the binding affinity between core targets and BAK. Results The results of CCK-8 and RTCA showed that BAK could inhibit the proliferation of nasopharyngeal carcinoma cells. The PI staining results showed that after 36-hour intervention with BAK in nasopharyngeal carcinoma 5-8F cells，BAK（30 μmol·L-1） increased the proportion of 5-8F cells in the S and G2/M phases，and the proportion of 6-10B cells in the G1 phase. BAK（40 μmol·L-1）increased the proportion of 5-8F cells in the sub G1 and S phases，and the proportion of 6-10B cells in the G2/M phase. The results of Hoechst 33342， JC-1， and Annexin V-FITC/PI dual fluorescence staining showed that BAK can cause cell condensation， fragmentation， and nuclear dissolution in 5-8F and 6-10B cells， reduce mitochondrial membrane potential，and increase cell apoptosis rate. The results of wound healing experiments and Transwell chamber method showed that BAK can reduce the migration and invasion rates of 5-8F and 6-10B cells. Western Blot results showed that BAK can reduce the expression levels of PCNA，Cyclin D3，XIAP，and Vimentin and N-cadherin proteins in 5-8F and 6-10B cells. Network pharmacology analysis showed that the MAPK signaling pathway in nasopharyngeal carcinoma is most affected by BAK. Molecular docking results showed that BAK has affinity with core target gene products VEGFA，PTGS2，EGFR，and TNF. Experimental verification showed that BAK can effectively reduce the expression levels of ERK1/2，p-ERK1/2，VEGFA，PTGS2，EGFR and TNF in 5-8F and 6- 10B cells. Conclusion BAK can inhibit the proliferation of nasopharyngeal carcinoma cells，induce apoptosis，inhibit cell migration and invasion. Its mechanism of action may be related to the impact on the MAPK signaling pathway and VEGFA，PTGS2，EGFR and TNF.

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国家自然科学基金项目（81973914）；湖南省自然科学基金项目（2023JJ30449，2023JJ30475）；湖南省教育厅科学研究项目 （21B0358，22A0254）；湖南省中医药管理局资助项目（D2022105）；湖南省卫健委项目（D202307017740）