【摘要】 目的 基于磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白（PI3K/Akt/mTOR）信号通路和多组学技术探讨异绿原酸A抗肝癌的作用机制。方法 分别考察0（对照组）、0.25、0.5 mg/mL的异绿原酸A干预人肝癌HepG2细胞48 h的侵袭率、迁移率，细胞中含DEP结构域的mTOR相互作用蛋白（DEPTOR）mRNA和mTOR、PI3K、第10号染色体缺失的磷酸酶及张力蛋白同源物（PTEN）蛋白表达量以及Akt蛋白磷酸化水平。采用液相色谱-串联质谱法进行代谢组学检测，筛选差异代谢物并进行京都基因与基因组百科全书（KEGG）通路富集分析；采用RNA测序进行转录组学检测，筛选差异表达基因并进行基因本体（GO）功能注释及KEGG通路富集分析。结果 与对照组比较，0.25、0.5 mg/mL异绿原酸A干预48 h能显著抑制HepG2细胞侵袭率和迁移率，显著上调DEPTOR mRNA和PTEN蛋白表达量，显著下调PI3K蛋白表达量和Akt蛋白磷酸化水平（0.25 mg/mL异绿原酸A除外）（P<0.05）。多组学分析共筛选出304个差异代谢物、212个差异表达基因；KEGG通路富集分析提示，异绿原酸A主要通过抑制PI3K/Akt信号通路，协同mTOR信号通路、铁死亡及磷酸戊糖途径、嘌呤/嘧啶代谢等代谢重编程过程，对HepG2细胞生长关键信号进行调控。结论 异绿原酸A的抗肝癌作用与阻断PI3K/Akt/mTOR信号通路异常激活相关，此外还可能与抑制磷酸戊糖途径及嘌呤/嘧啶代谢、诱导铁死亡等相关。更多还原

【Abstract】 OBJECTIVE To investigate the mechanism of isochlorogenic acid A against hepatocellular carcinoma based on the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR) signaling pathway and multiomics technology. METHODS The invasion rate and migration rate of human hepatocellular carcinoma HepG2 cells after 48 h of intervention with 0(control group), 0.25 and 0.5 mg/mL isochlorogenic acid A were examined; mRNA expression of DEP domaincontaining m TOR-interacting protein(DEPTOR), the protein expressions of mTOR, PI3K and phosphatase and tensin homologue deleted on chromosome ten(PTEN), as well as the phosphorylation level of Akt protein were determined in the cells. Metabolomics analysis was performed using liquid chromatography-tandem mass spectrometry, and differential metabolites were screened and subjected to Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis; transcriptomics monitoring was conducted by RNA sequencing, and differentially expressed genes were screened and subjected to gene ontology(GO) and KEGG pathway enrichment analyses. RESULTS Compared with the control group, intervention with 0.25 and 0.5 mg/mL isochlorogenic acid A for 48 h significantly inhibited the invasion rate and migration rate of HepG2 cells, significantly up-regulated the mRNA expression of DEPTOR and the protein expression of PTEN, and significantly down-regulated the protein expression of PI3K and the phosphorylation level of Akt protein(except for 0.25 mg/mL isochlorogenic acid A)(P<0.05). A total of 304 differential metabolites and 212 differentially expressed genes were screened by multi-omics analysis. KEGG pathway enrichment analysis suggested that isochlorogenic acid A regulated key signaling of HepG2 cell growth mainly by inhibiting the PI3K/Akt signaling pathway, synergizing with metabolic reprogramming such as m TOR signaling pathway, ferroptosis, pentose phosphate pathway and purine/pyrimidine metabolism. CONCLUSIONS The anti-hepatocellular carcinoma effect of isochlorogenic acid A is associated with the blockade of abnormal activation of the PI3K/Akt/mTOR signaling pathway. In addition, it may also be related to the inhibition of the pentose phosphate pathway and purine/pyrimidine metabolism, as well as the induction of ferroptosis,etc.更多还原