Beijing Institute of Hepatology Beijing You An Hospital Capital Medical University, Beijing, China.
Xuanwu Hospital, Capital Medical University, Beijing, China.
Naunyn Schmiedebergs Arch Pharmacol. 2023 Aug;396(8):1847-1856. doi: 10.1007/s00210-023-02428-2. Epub 2023 Mar 6.
In a previous study, our team found that ASPP2 overexpression increases the sensitivity of liver cancer cells to sorafenib. ASPP2 is an important target in the study of drug therapy for hepatocellular carcinoma. In this study, we demonstrated that ASPP2 altered the response of HepG2 cells to usnic acid (UA) by using mRNA sequencing and CyTOF. CCK8 assay was used to detect cytotoxicity of UA on HepG2 cells. Annexin V-RPE assay, TUNEL assay, and cleaved caspase 3 assay were performed to examine the apoptotic cell death induced by UA. Transcriptomic sequencing and a single-cell mass cytometry were used to analyze the dynamic response of HepG2shcon and HepG2shASPP2 cells to UA treatment. We have demonstrated that UA could inhibit proliferation in HepG2 cells in a concentration-dependent manner. Apoptotic cell death was significantly induced by UA in HepG2 cells, while knocking down ASPP2 could increase the resistance of HepG2 cells to UA. Data from mRNA-Seq indicated that knockout of ASPP2 in HepG2 cells affected cell proliferation, cycle, and metabolism. ASPP2 knockdown resulted in increased stemness and decreased apoptosis of HepG2 cells under the action of UA. CyTOF analysis confirmed the above results, ASPP2 knockdown increased oncoproteins in HepG2 cells and altered response patterns of HepG2 cells to UA. Our data suggested that the natural compound UA could inhibit liver cancer HepG2 cells; meanwhile, ASPP2 knockdown could affect response patterns of HepG2 cells to UA. The above results indicate that ASPP2 could be a research target in the chemoresistance of liver cancer.
在之前的研究中，我们的团队发现 ASPP2 过表达可增加肝癌细胞对索拉非尼的敏感性。ASPP2 是肝细胞癌药物治疗研究中的一个重要靶点。在本研究中，我们使用 mRNA 测序和 CyTOF 证实了 ASPP2 改变了 HepG2 细胞对乌头酸（UA）的反应。CCK8 检测试剂盒用于检测 UA 对 HepG2 细胞的细胞毒性。采用 Annexin V-RPE 检测试剂盒、TUNEL 检测试剂盒和 cleaved caspase 3 检测试剂盒检测 UA 诱导的细胞凋亡。采用转录组测序和单细胞质谱流式细胞术分析 HepG2shcon 和 HepG2shASPP2 细胞对 UA 处理的动态反应。结果表明，UA 可浓度依赖性地抑制 HepG2 细胞的增殖。UA 可显著诱导 HepG2 细胞发生凋亡，而敲低 ASPP2 可增加 HepG2 细胞对 UA 的耐药性。mRNA-Seq 数据表明，敲除 HepG2 细胞中的 ASPP2 会影响细胞的增殖、周期和代谢。ASPP2 敲低可增加 UA 作用下 HepG2 细胞的干性并减少细胞凋亡。CyTOF 分析证实了上述结果，ASPP2 敲低可增加 HepG2 细胞中的致癌蛋白并改变 HepG2 细胞对 UA 的反应模式。我们的数据表明，天然化合物 UA 可抑制肝癌 HepG2 细胞；同时，敲低 ASPP2 可影响 HepG2 细胞对 UA 的反应模式。上述结果表明，ASPP2 可能是肝癌化疗耐药性的研究靶点。
