肝细胞癌（HCC）是全球面临的重大健康挑战。化疗会导致肝癌细胞衰老。衰老的 HCC 细胞通过产生具有衰老相关分泌表型 (EV-SASP) 的细胞外囊泡，在抑制或促进癌症中发挥重要作用。在衰老过程中，EV-SASP 中的 miRNA 可以强烈上调，并且可以显着改变细胞的表型特征。 miRNA 微阵列分析显示，miRNA-146a-5p 在奥沙利铂和 H2O2 诱导的衰老 Huh7 细胞中高表达，RT-PCR 证实其在外泌体中显着上调。过表达miRNA-146a-5p的Huh7细胞的转录组测序结果表明，miRNA-146a-5p可以调节HCC细胞的糖酵解。随后，使用双荧光素酶测定来验证miRNA-146a-5p是否可以与IRF7相互作用以促进衰老。通过分析葡萄糖摄取、乳酸产生、耗氧率（OCR）和质子流出率（PER）的实验，确定了miRNA-146a-5p和IRF7在肝癌细胞有氧糖酵解中的关键功能。随后，通过荧光素酶报告基因检测证实了IRF7对关键糖酵解基因PFKL的调节作用。 Western blot实验结果表明，miR-146a-5p可以通过靶向IRF7激活CHK2和p53磷酸化蛋白，并上调p21蛋白。 miRNA-146a-5p的过度表达有效抑制HCC细胞的有氧糖酵解功能。此外，沉默 IRF7 可以有效抑制有氧糖酵解。 MiR-146a-5p。 MiR-146a-5p可以通过靶向IRF7激活CHK2磷酸化蛋白及其下游蛋白p53的磷酸化，激活的p53上调p21的表达。我们的研究表明，衰老HCC细胞产生的外泌体miRNA-146a-5p可以通过抑制有氧糖酵解来抑制HCC细胞增殖，并通过靶向IRF7激活CHK2/p53/p21信号通路来促进HCC细胞衰老。©作者。

Hepatocellular carcinoma (HCC) is a major global health challenge. Chemotherapy can cause HCC cells to become senescent. Senescent HCC cells play an important role in inhibiting or promoting cancer by producing extracellular vesicles with a senescence-associated secretory phenotype (EV-SASP). miRNA can be strongly upregulated in EV-SASP during the aging process and can substantially alter the phenotypic characteristics of cells. MiRNA microarray analysis revealed that miRNA-146a-5p was highly expressed in oxaliplatin- and H2O2-induced senescent Huh7 cells, and RT‒PCR confirmed its significant upregulation in exosomes. The transcriptome sequencing results of Huh7 cells overexpressing miRNA-146a-5p suggested that miRNA-146a-5p could regulate HCC cell glycolysis. Subsequently, a dual luciferase assay was used to verify whether miRNA-146a-5p can interact with IRF7 to promote aging. The key functions of miRNA-146a-5p and IRF7 in aerobic glycolysis in liver cancer cells were determined through experiments analyzing glucose uptake, lactate production, the oxygen consumption rate (OCR) and the proton efflux rate (PER). Subsequently, the regulatory effect of IRF7 on the key glycolytic gene PFKL was confirmed through luciferase reporter assays. The western blot experiment results showed that miR-146a-5p can activate CHK2 and p53 phosphorylated proteins by targeting IRF7, and upregulate p21 protein. Overexpression of miRNA-146a-5p effectively inhibited the aerobic glycolytic function of HCC cells. Moreover, silencing IRF7 effectively inhibited aerobic glycolysis. MiR-146a-5p. MiR-146a-5p can activate the phosphorylation of CHK2 phosphorylation protein and its downstream protein p53 by targeting IRF7, and the activated p53 upregulates the expression of p21. Our study revealed that exosomal miRNA-146a-5p produced by aging HCC cells, can inhibit HCC cell proliferation through inhibiting aerobic glycolysis and promote HCC cell aging by activating CHK2/p53/p21 signaling way by targeting IRF7.© The author(s).