益肾骨康方对肺癌A549细胞形态及转录组基因表达的影响  

Effect of Yishen Gukang Prescription on Morphology and Transcriptome Gene Expression of Lung Cancer A549 Cells

作　　者：何理玉 杨梦霞 芦殿荣[1] 冯利 HE Liyu;YANG Mengxia;LU Dianrong;FENG Li(Wangjing Hospital,China Academy of Chinese Medical Sciences,Beijing 100102,China;National Cancer Center/National Clinical Research Center for Cancer,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China)

机构地区：[1]中国中医科学院望京医院,北京100102 [2]国家癌症中心/国家肿瘤临床医学研究中心/中国医学科学院北京协和医学院肿瘤医院,北京100021

出　　处：《世界中医药》2022年第15期2162-2166,共5页World Chinese Medicine

基　　金：国家自然科学基金青年基金项目(81603598)——基于OPG/RANKL/RANK轴及其关键分子探讨扶正解毒化瘀中药益肾骨康方防治骨转移癌的作用机制。

摘　　要：目的:通过益肾骨康方作用后肺癌A549细胞基因的差异表达情况及进行功能分析,探讨益肾骨康方作用于肺癌的分子作用机制奠定实验基础。方法:实验分为对照组和益肾骨康方组,中药作用于A549细胞24 h后,利用扫描电镜观察A549细胞表面形态的变化;利用转录组基因测序,筛选差异表达基因,并通过基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析对差异表达基因进行功能富集分析。利用Cytoscape 3.6.0软件构建差异表达基因蛋白质-蛋白质相互作用(PPI)网络。结果:扫描电镜结果显示对照组细胞形态正常,饱满,表面有膜皱,丝状伪足。益肾骨康方组可见细胞表面膜皱显著减少,丝状伪足断裂,凋亡小体形成。差异基因筛选|log2FoldChange|>1的差异表达基因有440个,其中下调基因有136个,上调基因有304个。其中与癌症密切相关的基因为SOX2,在给药组中表达下调。在GO功能富集过程中共得到307条结果,主要涉及生物过程(237项)、细胞组分(56项)和分子功能(14项)。KEGG通路分析,结果显示与益肾骨康方抑制肺癌A549细胞的作用相关机制主要涉及到细胞凋亡方面。通过构建PPI网络,找到与中药干预肺癌增殖和转移相关的2个节点度较高的基因:MMP9和CCL2。与对照组比较益肾骨康方下调了MMP9和CCL2基因的表达。结论:益肾骨康方的抗肿瘤作用机制可能是通过下调SOX2、MMP9和CCL2基因的表达,从而抑制了肺癌A549细胞的增殖和转移能力。Objective:To explore the molecular mechanism of Yishen Gukang Prescription on lung cancer by analyzing the differential expression and functions of A549 cell gene after Yishen Gukang Prescription intervention.Methods:The experiment was divided into the control group and the Yishen Gukang Prescription group.After treatment with Yishen Gukang Prescription for 24 h,the changes of surface morphology of A549 cells were observed by scanning electron microscope.Transcriptome gene sequencing was conducted to screen differentially expressed genes,and functional enrichment analysis of the differentially expressed genes was carried out through Gene Ontology(GO)and Kyoto Gene and Genome Encyclopedia(KEGG),and Cytoscape3.6.0 was used to construct the protein-protein interaction(PPI)network.Results:The scanning electron microscopy indicated that the cells in the control group were normal,plump,and wrinkled with filamentous shape.In the Yishen Gukang Prescription group,membrane wrinkles on cell surface were significantly reduced,with damaged filamentous shape and apoptotic bodies formed.There were a total of 440 differentially expressed genes with|log2 FoldChange|>1,among which 304 were up-regulated and 136 were down-regulated.One gene closely related to cancer,SOX2,was down-regulated in the Yishu Gukang Prescription group.A total of 307 items were obtained in the GO analysis,mainly involving biological processes(237 items),cellular components(56 items)and molecular functions(14 items).KEGG pathway analysis revealed that the mechanism associated with Yishen Gukang Prescription in inhibiting lung cancer A549 cells was mainly involved in apoptosis.By the construction of PPI network,two genes with high node degree related to the intervention of Yishen Gukang Prescription on lung cancer proliferation and metastasis were found:MMP9 and CCL2.Compared with the condition in the control group,Yishen Gukang Prescription down-regulated the expression of MMP9 and CCL2.Conclusion:The antitumor mechanism of Yishen Gukang Prescription might b

关 键 词：中药 肺癌 A549细胞 益肾骨康方 扫描电镜 转录组基因测序 生物信息学 核心基因 

分 类 号：R289.5[医药卫生—方剂学] R734.2[医药卫生—中药学]