Chen Q, Shang S, Lu H, Li S, Sun Z, Fan X, Qi Z
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Wannan Medical College, Wuhu 241002, China.
Anhui Provincial Key Laboratory of Active Biological Macro-molecules, Wannan Medical College, Wuhu 241002, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Jul 20;44(7):1327-1335. doi: 10.12122/j.issn.1673-4254.2024.07.12.
To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma (HCC) cell proliferation and migration.
HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4, SLC7A11, LC3, P62 and phosphorylation of Akt/mTOR using Western blotting. The effects LY294002 and Rapamycin (the inhibitor and activator of autophagy, respectively) on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays. The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes. GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.
Calenduloside E obviously inhibited the viability of HCC cells. GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines, and their expression levels were negatively correlated with the patients'survival. In HCC cell lines, calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins, activated the Akt-mTOR pathway, and enhanced the expression of LC3 Ⅱ. The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002. Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells, while activating this pathway produced the opposite effect.
Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.
探讨金盏花苷E抑制肝细胞癌（HCC）细胞增殖和迁移的分子机制。
用CCK-8法检测经金盏花苷E处理的HCC细胞系HepG2和Huh7的细胞活力变化，用蛋白质免疫印迹法检测谷胱甘肽过氧化物酶4（GPX4）、溶质载体家族7成员11（SLC7A11）、微管相关蛋白轻链3（LC3）、p62的表达以及蛋白激酶B（Akt）/哺乳动物雷帕霉素靶蛋白（mTOR）的磷酸化水平。用5-乙炔基-2'-脱氧尿苷（EdU）法和Transwell法评估自噬抑制剂LY294002和自噬激活剂雷帕霉素对经金盏花苷E处理的HCC细胞增殖和迁移的影响。利用癌症基因组图谱（TCGA）数据库探究HCC组织和正常肝组织中GPX4和SLC7A11的表达水平及其与患者生存结局的相关性。用逆转录-定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测HCC细胞和正常肝细胞中GPX4和SLC7A11的表达。
金盏花苷E明显抑制HCC细胞活力。GPX4和SLC7A11在HCC组织和细胞系中高表达，其表达水平与患者生存率呈负相关。在HCC细胞系中，金盏花苷E显著抑制GPX4和SLC7A11蛋白的表达，激活Akt-mTOR通路，并增强LC3Ⅱ的表达。雷帕霉素显著增强金盏花苷E对GPX4和SLC7A11表达的抑制作用，而LY294002则减弱该抑制作用。抑制自噬通路明显减弱金盏花苷E对HCC细胞增殖和迁移的抑制作用，而激活该通路则产生相反的效果。
金盏花苷E通过自噬通路下调GPX4和SLC7A11的表达来抑制HCC细胞的增殖和迁移。
