麦门冬汤合千金苇茎汤对小鼠Lewis原位肺癌lncRNA,mRNA表达谱的影响

Effect of Maimendong Tang and Qianjin Weijingtang on Expression Profiles of lncRNA and mRNA in Mouse Orthotopic Lewis Lung Carcinoma

目的:通过对Lewis肺癌原位移植小鼠肿瘤组织长链非编码RNA(long noncoding RNA,lncRNA)和mRNA表达谱的检测,探讨麦门冬汤合千金苇茎汤(金方)抗肺癌的作用机制。方法:C57BL/6小鼠随机分为正常组、模型组、金方组(20 g·kg-1·d-1),构建小鼠Lewis原位肺癌模型成功后,次日用金方灌胃开始治疗。应用基因芯片技术,检测与金方抗肺癌作用密切相关的差异lncRNA,mRNA,进行聚类分析,并利用t检验和差异表达的倍数变化,筛选出关键lncRNA,mRNA。运用生物信息学方法预测差异lncRNA调控的靶基因,并对其进行功能和通路分析。应用病理组织学技术,检测各组小鼠肿瘤组织光镜下的差异表现。结果:lncRNA,mRNA芯片杂交结果显示,金方调控了887个lncRNA差异表达,其中上调442个,下调445个(P<0.05)。差异表达的mRNA共610个,其中上调376个,下调234个(P<0.05)。使用GO分析方法,发现下调或上调的靶基因主要参与了细胞代谢调控、信号通路调控等生物学过程(biological process,BP)。靶基因分子功能(molecular function,MF)分析显示,下调或上调的靶基因主要具有DNA结合、蛋白质结合、受体结合和转录因子活性等功能。信号通路预测显示金方可调控Janus激酶/信号转导与转录激活因子(JAK/STAT)等与肺癌的发生发展密切相关的信号通路。病理组织学检查结果证实,与模型组比较,金方能明显改善小鼠肺组织的病理改变。结论:金方可能通过调控多种lncRNA和mRNA的表达及下游相关信号通路来发挥其抗肺癌的作用。

Objective:To explore the anti-lung cancer mechanism of Maimendong Tang and Qianjin Weijingtang(Jin Fang)by detecting the expression profiles of long noncoding RNA(lncRNA)and mRNA in mice tumor tissues of orthotopic Lewis lung cancer model.Method:C57 BL/6 mice were randomly divided into normal group,model group and Jin Fang group(20 g·kg-1·d-1).After successful establishment of Lewis lung cancer model in situ in mice,Jin Fang was given orally the next day after treatment.Using gene chip technology,differential lncRNA and mRNA closely related with Jin Fang’s anti-lung cancer effect were detected,and cluster analysis was performed.The key lncRNA and mRNA were screened out by t-test and fold change of differential expression.Bioinformatic methods were used to predict target genes regulated by differential lncRNA,and functional and pathway analysis was performed.The histopathological technique was used to detect the differences in the tumor tissue of each group under light microscope.Result:lncRNA and mRNA chip hybridization results showed that Jin Fang regulated differential expressions of 887 lncRNA,in which 442 were up-regulated and 445 were down-regulated(P<0.05).There were 610 differential mRNA expressions,in which 376 were up-regulated and 234 were down-regulated(P<0.05).GO analysis showed that down-regulated or up-regulated target genes were mainly involved in biological processes(BP),such as cell metabolism regulation and signal pathway regulation.Molecular function(MF)analysis showed such functions DNA binding,protein binding,receptor binding and transcription factor activity in down-regulated or up-regulated target genes.Target genes were involved in multiple biological processes,cellular components and molecular functions.Signal pathway predictions indicated that Jin Fang can regulate the Janus kinase/signal transducer and activator of tran-ions(JAK/STAT)and other signaling pathways closely related to the development of lung cancer.The results of histopathological examination confirmed that Jin Fang could significantly improve the pathological changes of lung tissues in the mice compared with the model group.Conclusion:Jin Fang may exert its anti-lung cancer effect by regulating the expressions of multiple lncRNAs and mRNAs,and down-regulating related signaling pathways.