乌梅丸对乳腺癌肺转移及转移微环境核心信号转导分子的影响  

Effect of Wumei Pill(乌梅丸)on Lung Metastasis and Core Signal Transduction Molecules in Metastatic Microenvironment of Breast Cancer

作　　者：王萍[1] 任美玲 吴紫葶 李锦超 杨海燕[1] WANG Ping;REN Meiling;WU Ziting;LI Jinchao;YANG Haiyan(Jiangxi University of Chinese Medicine,Nanchang 330004,Jiangxi,China)

机构地区：[1]江西中医药大学,江西南昌330004

出　　处：《中华中医药学刊》2023年第12期1-8,I0021-I0023,共11页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81703980);江西省自然科学基金项目(20202BABL206129);江西省教育厅科技项目(209804);江西省中医药管理局中医证候基础重点研究室项目(赣中医药科教字[2022]8号-4);江西中医药大学科技创新团队发展项目(CXTD22016)。

摘　　要：目的通过与乳腺癌常用化疗药物卡培他滨的比较,分析乌梅丸对小鼠乳腺癌肺转移微环境基因表达的影响以及对微环境中核心信号转导分子的影响。方法采用luciferase稳转小鼠乳腺癌细胞4T1接种于SPF级Balb/c雌性小鼠的第2对乳腺脂肪垫内建立乳腺癌动物模型,分为药物观察组(临床折算剂量的乌梅丸)、药物对照组(临床折算剂量的卡培他滨)和空白对照组。利用生物发光活体成像技术观察乳腺癌在接种原位及肺部的生长转移情况。利用Agilent表达谱芯片检测了乳腺癌肺部转移微环境中的基因表达情况,筛选出差异表达基因并进行生物信息学分析。比较乌梅丸与卡培他滨干预后肺部转移微环境中基因表达的差异。并进一步分析乌梅丸对肺部转移微环境中核心信号转导分子的影响。利用STRING在线分析系统等分析获得核心信号转导分子。最后通过RT-PCR、免疫组化及蛋白质免疫印迹方法检测了这些信号转导分子的表达情况。结果随着时间的推移各组小鼠乳腺癌接种处荧光面积逐渐增大,显示肿瘤体积在不断增加;在第30天时间点上空白对照组的荧光分布分散,已超出原发接种灶附近,表示肿瘤存在明显转移情况,较药物观察组与药物对照组明显。药物观察组、药物对照组分别与空白对照组比较,共有23个共同调控基因,但药物观察组所影响的基因明显少于药物对照组,二者的基因本体(Gene ontology,GO)和京都基因和基本组数据库(Kyoto encyclopedia of genes and genomes,KEGG)pathway无共同条目。通过分析药物观察组与空白对照组比较获得的123个差异表达基因的信号转导网络,首要推荐核心基因为AGT、CCL20、CXCL9、CXCL10、HTR1B。RT-PCR结果显示药物观察组AGT、CCL20、CXCL9、CXCL10、HTR1B的表达与空白对照组比较均降低,差异具有统计学意义(P<0.01)。免疫组化结果显示药物观察组的核心基因呈�Objective By comparing with capecitabine,a commonly used chemotherapy drug for breast cancer,the effect of Wumei Pill(乌梅丸)on gene expression in lung metastasis microenvironment and core signal transduction molecules in microen⁃vironment of breast cancer in mice was analyzed.Methods Breast cancer cell 4T1 was inoculated into the second pair of breast fat pads of SPF balb/c female mice to establish an animal model of breast cancer.The animal model was divided into drug observa⁃tion group[Wumei Pill with clinical converted dose),drug control group(capecitabine with clinical converted dose)and blank control group.Bioluminescence in vivo imaging technique was used to observe the growth and metastasis of breast cancer in situ and lung.The gene expression in the microenvironment of lung metastasis of breast cancer was detected by Agilent expression mi⁃croarray,and the differentially expressed genes were screened and analyzed by bioinformatics.The difference of gene expression in lung metastasis microenvironment between Wumei Pill and capecitabine after intervention.And it further analyzed the effect of Wumei Pill on the core signal transduction molecules in the lung metastasis microenvironment.The core signal transduction mole⁃cules were obtained by using string online analysis system.Finally,the expressions of these signal transduction molecules were detected by RT-PCR,immunohistochemistry and Western blot.Results With the time passing by,the fluorescent area of breast cancer inoculation site of mice in each group gradually increased,indicating that the tumor volume was increasing.On the 30th day,the fluorescence distribution of the blank control group was scattered,which had exceeded the vicinity of the primary vaccina⁃tion focus,indicating that there was obvious metastasis of the tumor,which was more obvious than that of the drug observation group and the drug control group.Compared with the blank control group,the drug observation group and the drug control group had a total of 23 common regulatory g

关 键 词：乌梅丸 卡培他滨 乳腺癌 转移微环境 信号转导 

分 类 号：R255.2[医药卫生—中医内科学]