摘要
目的:通过观察乌梅丸干预后对乳腺癌小鼠肺部微环境基因表达的影响以及相关的生物信息学分析,探寻乌梅丸调控肺部微环境基因表达的特征,以评价乌梅丸改善肺部微环境抑制乳腺癌肺转移的作用机制。方法:以荧光素酶转染的乳腺癌4T1细胞的原位移植构建乳腺癌模型,观察原位移植瘤体积以及生物发光活体成像法观察移植瘤生长转移情况;采用直接观察法和HE染色观察肺部整体及微观情况;以表达谱芯片分析乌梅丸16.4 g/kg组、模型对照组和正常对照组肺组织的基因表达情况,筛选乳腺癌肺部微环境改变基因以及乌梅丸调控的肺部微环境基因;生物信息学方法进行基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书(KEGG)数据库分析乌梅丸调控的肺部微环境基因的特点;RT-PCR法、免疫组化法和蛋白质印迹法检测7个挑选的乌梅丸调控的肺部微环境基因的表达。结果:与正常对照组比较,模型对照组肺部外观和HE染色均提示肺部的病理改变,且肺部微环境有2553个基因表达有差异,GO共有443条富集项,其中生物学过程(BP)计292条,细胞组件(CC)计80条;分子功能(MF)计71条;KEGG分析共有38条富集项,剔除非相关性的疾病信号通路,还有20条信号通路。与模型对照组比较,卡培他滨0.36 g/kg组和乌梅丸16.4 g/kg组均能改善肺部外观和病理改变,并抑制原位移植瘤的生长和转移,卡培他滨0.36 g/kg组更为明显;与模型对照组比较,乌梅丸16.4 g/kg组肺部微环境基因有123个基因表达有差异,其中64个基因上调,59个基因下调;GO分析有3条富集项,其中BP共计1条,CC共计0条,MF共计2条;KEGG分析无条目同时满足P≤0.05和FDR≤0.05的筛选条件。与模型对照组比较,采用RT-PCR方法分析了6个感兴趣的基因,其中Npr3、Igll1、Rasl11a、Arntl 4个基因的表达在乌梅丸干预后下调,Per3和Dbp 2个基因在乌梅丸干预后表达上调(P<0.
Objective:To study the characteristics of Wumeiwan(乌梅丸)in regulating gene expression of lung microenvironment of the mice suffering from breast cancer,so as to decipher the mechanism of Wumeiwan in improving lung microenvironment and inhibiting lung metastasis of breast cancer.Methods:The model of breast cancer was established by orthotopic transplantation with the breast cancer 4T1 cells transfected with luciferase.The volumes of orthotopic transplantation tumors were observed,and the tumor growth and metastasis were evaluated by the bioluminescent imaging method.The whole and microscopic conditions of the lung were observed directly and by HE staining.The expression microarray was used to determine the gene expression of the lung tissue in the 16.4 g/kg Wumeiwan,model control,and normal control groups.The lung microenvironment genes regulated by breast cancer and Wumeiwan were screened out.Gene ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were carried out to analyze the lung microenvironment genes regulated by Wumeiwan.RT-PCR,immunohistochemistry,and Western blotting were employed to examine the expression of six genes involved in the regulation of lung microenvironment by Wumeiwan.Results:Compared with normal control group,the modeling caused pathological changes and differential expression of 2553 genes in the lung microenvironment of breast cancer.These differentially expressed genes(DEGs)were annotated into 443 GO terms(292 biological process terms,80 cell component terms,and 71 molecular function terms).In addition,these genes were enriched in 38 KEGG pathways,in which 20 signaling pathways remained after excluding unrelated disease signaling pathways.Compared with the model control group,0.36 g/kg xeloda and 16.4 g/kg Wumeiwan alleviated the pathological changes in the lung tissue and inhibited the growing and transferring of orthotopic transplantation tumor,and the former 0.36 g/kg xeloda showed stronger effect.Compared with the model control group,16.4
作者
王萍
杨海燕
吴紫葶
任美玲
李锦超
WANG Ping;YANG Haiyan;WU Ziting;REN Meiling;LI Jinchao(College of Chinese Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2023年第11期30-39,共10页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金(编号:81703980)
江西省自然科学基金(编号:20202BABL206129)
江西省教育厅科技项目(编号:209804)
江西中医药大学科技创新团队发展项目(编号:CXTD22016)
江西省中医药管理局中医证候基础重点研究室(赣中医药科教字[2022]8号-4)。
关键词
乌梅丸
乳腺癌
转移灶微环境
表达谱芯片
生物信息学
Wumeiwan(乌梅丸)
Breast cancer
Metastasis microenvironment
Expression microarray
Bioinformatics
