基于转录组学探讨白花蛇舌草总黄酮抗肺癌A549细胞生长机制

Transcriptomic study of total flavonoids of Hedyotis diffusa Willd.against the growth of lung cancer cells

目的 利用转录组学技术揭示白花蛇舌草总黄酮提取物抑制肺癌细胞生长的作用机制。方法 首先制备白花蛇舌草总提取物和总黄酮提取物,采用CCK8法检测不同剂量的白花蛇舌草总提取物和总黄酮(50、100、200 mg·L^(-1))对肺癌细胞株A549细胞增殖活性的影响。然后,选择最佳剂量的总黄酮干预肺癌细胞,采用CCK8法、划痕实验、Transwell侵袭实验检测白花蛇舌草总黄酮对肺癌细胞增殖、迁移和侵袭能力的影响,并进行转录组测序。最后,对转录组数据分别进行差异分析、功能富集分析、蛋白质-蛋白质相互作用(PPI)分析。结果 白花蛇舌草总提取物和总黄酮提取物均能显著抑制肺癌细胞增殖活性,且总黄酮作用优于总提取物。转录组学分析共筛选出1 134个差异表达基因,这些基因富集于MAPK信号通路、WNT信号通路、转化生长因子β信号通路、p53信号通路等多条癌症相关信号通路。从差异基因的PPI网络中筛选得到10个关键基因,分别为TTK、NUSAP1、CDCA8、KIF2C、MELK、CCNB2、CCNB1、KIF20A、BUB1、CEP55。结论 白花蛇舌草总黄酮成分抗肺癌细胞生长的作用机制可能涉及MAPK信号通路、WNT信号通路、转化生长因子β信号通路、p53信号通路等癌症相关信号通路,并且可能直接作用于TTK、NUSAP1、CDCA8、KIF2C、MELK、CCNB2、CCNB1、KIF20A、BUB1、CEP55等癌基因。

Objective To revealing the mechanism total flavonoid extract of Hedyotis diffusa Willd.in inhibiting the growth of lung cancer cells using transcriptomic techniques.Methods The total extract and total flavonoid extract of Hedyotis diffusa Willd.were firstly prepared.The effects of different doses of total extract and total flavonoids of Hedyotis diffusa Willd.(50,100 and 200 mg·L^(-1))on the proliferation activity of A549 cells were examined by CCK8 method.Then,transcriptomic techniques were applied to detect the gene expression patterns in the blank control group and the experimental group(100 mg·L^(-1) total flavonoids of Hedyotis diffusa Willd.intervention).Finally,the transcriptomic data were subjected to differential analysis,functional enrichment analysis,and protein-protein interaction(PPI)analysis,respectively.Results Both total extract and total flavonoid extract of Hedyotis diffusa Willd.significantly inhibited the proliferation activity of lung cancer cells,and the total flavonoid effect was superior to the total extract.Transcriptomic analysis screened a total of 1134 differentially expressed genes,which were enriched in several cancer-related signaling pathways such as MAPK signaling pathway,WNT signaling pathway,transforming growth factorβsignaling pathway and p53 signaling pathway.Ten key genes were screened from the PPI network of differential genes,namely TTK,NUSAP1,CDCA8,KIF2C,MELK,CCNB2,CCNB1,KIF20A,BUB1,and CEP55.Conclusion The mechanism of total flavonoid components of Hedyotis diffusa Willd.against lung cancer cell growth may involve cancer-related signaling pathways such as MAPK signaling pathway,WNT signaling pathway,transforming growth factorβsignaling pathway,p53 signaling pathway,and may act directly on oncogenes such as TTK,NUSAP1,CDCA8,KIF2C,MELK,CCNB2,CCNB1,KIF20A,BUB1,CEP55,etc.