高通量测序筛选丹参活性成分调控人肺腺癌A549细胞lncRNA-mRNA相关性研究  

Study on the correlation between the active components of Salviea Miltiorrhizae Radix et Rhizoma screened by high-throughput sequencing and the regulation of lncRNA-mRNA in human lung adenocarcinoma A549 cells

作　　者：郑琦[1] 贺妍 薛超[1] 朱广辉 王学谦[1] 侯炜[1] Zheng Qi;He Yan;Xue Chao;Zhu Guanghui;Wang Xueqian;Hou Wei(Oncology Department,Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China;Graduate School of Chinese Academy of Agriculture Sciences,Beijing 100081,China)

机构地区：[1]中国中医科学院广安门医院肿瘤科,北京100053 [2]中国农业科学院研究生院,北京100081

出　　处：《国际中医中药杂志》2024年第4期479-484,共6页International Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82104656、82004179);中央级公益性科研院所基本科研业务费专项资金(ZZ15-YQ-024)。

摘　　要：目的筛选丹参活性成分隐丹参酮、丹参酮ⅡA对人肺腺癌A549细胞相关差异表达的长链非编码RNA(lncRNA)、信使RNA(mRNA),构建基因调控网络,从基因水平探索丹参活性成分调控人肺腺癌A549细胞的物质基础和作用机制。方法培养A549细胞,通过细胞增殖实验确认隐丹参酮、丹参酮ⅡA的有效剂量。将A549细胞按随机数字表法分为空白对照组、隐丹参酮组及丹参酮ⅡA组,干预24 h后,流式细胞仪检测细胞周期,并使用高通量测序技术检测各组A549细胞中lncRNA、mRNA表达情况。通过分析隐丹参酮、丹参酮ⅡA作用的相关差异基因表达谱,对差异基因进行GO功能和KEGG通路富集分析。结果细胞周期结果显示,隐丹参酮组、丹参酮ⅡA组G0/G1期细胞比例升高(P<0.01),S期细胞比例降低(P<0.01),隐丹参酮组G2/M期比例降低(P<0.01)。高通量测序结果发现,隐丹参酮能够上调4698个lncRNA,下调1557个lncRNA,上调4810个mRNA,下调5644个mRNA。丹参酮ⅡA能够上调1348个lncRNA,下调1299个lncRNA,上调4646个mRNA,下调4741个mRNA。将有差异的lncRNA和mRNA进行功能与通路富集分析显示,隐丹参酮、丹参酮ⅡA差异表达基因主要与细胞周期、自噬、AMPK信号通路、FoxO信号通路、EGFR信号通路等相关。GAS5可能是隐丹参酮和丹参酮ⅡA起到抑制作用的靶点之一。结论隐丹参酮、丹参酮ⅡA对A549细胞有一定的抑制作用,存在lncRNA-mRNA差异表达基因谱,其与细胞周期及信号通路关系密切。Objective Study on the correlation between the active components of Salviea Miltiorrhizae Radix et Rhizoma screened by high-throughput sequencing and the regulation of lncRNA-mRNA in human lung adenocarcinoma A549 cells.Methods A549 cells were cultured,and the IC50 dose of cryptotanshinone and tanshinoneⅡA was confirmed according to the cell proliferation experiment.A549 cells were randomly divided into blank control group,cryptotanshinone group,and tanshinone IIA group using a random number table method.After 24 hours of intervention,the cell cycle was detected by flow cytometry.High-throughput sequencing technique was used to detect the expressions of lncRNA and mRNA in A549 cells in intervention group and non-intervention group.By analyzing the expression profiles of differential genes related to cryptotanshinone and tanshinoneⅡA,the obtained differential genes were analyzed by GO and KEGG.Results The cell cycle results showed that the proportion of G0/G1 phase cells in cryptotanshinone and TanshinoneⅡA was increased(P<0.01),the proportion of S phase cells was decreased(P<0.01),and the proportion of G2/M phase cells in cryptotanshinone was decreased(P<0.01).The results of high-throughput screening showed that cryptotanshinone could up-regulate 4698 lncRNA,down-regulate 1557 lncRNA,up-regulate 4810 mRNA and down-regulate 5644 mRNA.TanshinoneⅡA could up-regulate 1348 lncRNA,down-regulate 1299 lncRNA,up-regulate 4646 mRNA and down-regulate 4741 mRNA.The function and pathway enrichment analysis of differential lncRNA and mRNA showed that the differentially expressed genes of cryptotanshinone and tanshinoneⅡA were mainly related to cell cycle,autophagy,AMPK signaling pathway,FoxO signaling pathway and EGFR signaling pathway.GAS5 may be one of the targets for the inhibitory effects of cryptotanshinone and tanshinoneⅡA.Conclusion Cryptotanshinone and tanshinoneⅡA have certain inhibitory effects on A549 cells,and there are differentially expressed genes of lncRNA-mRNA,which are closely related to cell cy

关 键 词：隐丹参酮 丹参酮ⅡA 肺癌 lncRNA测序 转录组测序 细胞周期 

分 类 号：R285[医药卫生—中药学]