槲皮素的作用靶点预测及其对肝内胆管癌细胞RBE增殖、迁移及侵袭的影响机制  

Prediction of quercetin s target of action and its mechanism of influence on proliferation,migration and invasion of intrahepatic cholangiocarcinoma cell RBE

作　　者：夏果毅 李巨仕 张鸿渐 曾稳盈 姜志胜 郑核 XIA Guoyi;LI Jushi;ZHANG Hongjian;ZENG Wenying;JIANG Zhisheng;ZHENG He(Department of Hepatobiliary Surgery,Shaoyang Central Hospital,Affiliated Shaoyang Hospital,University of South China,Shaoyang 422000,Hunan,China;Cardiovascular Disease Research Institute,Hengyang Medical School,University of South China,Hengyang 421001,Hunan,China)

机构地区：[1]邵阳市中心医院,南华大学附属邵阳医院肝胆外科,湖南邵阳422000 [2]南华大学衡阳医学院心血管疾病研究所,湖南衡阳421001

出　　处：《中南医学科学杂志》2025年第1期17-22,共6页Medical Science Journal of Central South China

基　　金：国家自然科学基金资助项目(91839103,81670429);湖南省卫生健康委科研计划项目资助(202204013436);邵阳市科技计划项目经费资助(2022GZ4161)。

摘　　要：目的预测槲皮素治疗肝内胆管癌的作用靶点,并探讨其对肝内胆管癌细胞RBE增殖、迁移及侵袭的影响机制。方法GeneCards数据库获得肝内胆管癌相关基因,SwissTargetPrediction数据库预测槲皮素的潜在作用靶点。通过STRING数据库构建蛋白互作网络进行可视化分析。利用GO和KEGG分析相关基因富集通路。通过体外实验,将肝内胆管癌细胞RBE分为对照组、槲皮素组。CCK-8法、克隆形成实验、划痕实验、Transwell实验检测槲皮素对RBE细胞增殖、迁移和侵袭的影响。槲皮素处理RBE细胞后收集RNA进行测序、流式细胞仪检测活性氧(ROS)水平,Western blotting检测磷酸化(p)-P65/P65水平。结果预测到槲皮素治疗肝内胆管癌的潜在作用靶点46个,主要富集于PI3K-AKT信号通路、氧化应激反应、含磷基团的转移酶复合体和蛋白丝氨酸/苏氨酸/酪氨酸激酶活性信号通路等。与对照组相比,槲皮素组抑制了RBE细胞的增殖、迁移和侵袭;RNA测序结果显示,差异基因富集于肿瘤坏死因子-α、核因子(NF)-κB、ROS、炎症反应、铁死亡、甾体激素生物合成和D-苏醛糖1-脱氢酶活化等信号通路;流式细胞术和Western blotting结果显示,槲皮素能升高ROS,并降低p-P65的表达。结论NF-κB是槲皮素抑制肝内胆管癌细胞增殖、迁移、侵袭的关键靶点,其分子机制可能是槲皮素促进ROS水平从而抑制NF-κB信号通路。Aim To predict the target of quercetin in the treatment of intrahepatic cholangiocarcinoma,and analyze its mechanism of influence on the proliferation,migration,and invasion of intrahepatic cholangiocarcinoma cell RBE.Methods Genes related to intrahepatic cholangiocarcinoma were obtained by using the GeneCards database.The SwissTargetPrediction database was utilized to predict potential target of quercetin.A protein-protein interaction network was constructed and visualized by using the STRING database.Pathway enrichment analysis of the relevant genes was conducted by using the GO and KEGG.In vitro experiments were performed by dividing intrahepatic cholangiocarcinoma RBE cells into a control group and a quercetin-treated group.The effects of quercetin on RBE cell proliferation,migration,and invasion were assessed by using CCK-8 assays,clone formation assays,wound healing assays,and Transwell assays.After treating RBE cells with quercetin,RNA was collected for sequencing,reactive oxygen species(ROS)levels were measured by using flow cytometry,and the levels of phosphorylation(p)-P65/P65 were detected by Western blotting.Results Forty-six potential targets for quercetin in the treatment of intrahepatic cholangiocarcinoma have been predicted,primarily enriched in the PI3K-AKT signaling pathway,response to oxidative stress,transferase complex,transferring phosphorus-containing groups,and protein serine/threonine/tyrosine kinase activity signaling pathways.Compared with the control group,the quercetin group inhibited the proliferation,migration,and invasion of RBE cells.RNA sequencing results indicated that differentially expressed genes were enriched in signaling pathways such as tumor necrosis factor(TNF-α),nuclear factor(NF)-κB,ROS,inflammatory response,ferroptosis,steroid hormone biosynthesis,and D-threo-aldose 1-dehydrogenase activity.Flow cytometry and Western blotting results demonstrated that quercetin could increase ROS levels and decrease the expression of p-P65.Conclusion NF-κB was found to be a key tar

关 键 词：槲皮素 肝内胆管癌 治疗靶点 RBE 增殖 迁移 侵袭 

分 类 号：R735.8[医药卫生—肿瘤]