2',4'-二羟基-3'-甲基-3-甲氧基查耳酮上调p21抑制人肝癌HepG2细胞的生长
2',4'-dihydroxy-3'-methyl-3-methoxychalcone inhibits growth of human hepatocellular carcinoma HepG2 cells through up-regulation of p21
王龙燕 1张云封 1王柱国 1谭鹏 1魏雪娇 1李军 2胡仲冬2
作者信息
- 1. 北京中医药大学中药学院,北京 100029;北京中医药大学北京中医药研究院中药现代研究中心,北京 100029
- 2. 北京中医药大学北京中医药研究院中药现代研究中心,北京 100029
- 折叠
摘要
目的 探讨2',4'-二羟基-3'-甲基-3-甲氧基查耳酮(C20)对人肝癌HepG2细胞的体外抗肿瘤作用及其潜在的作用机制.方法 通过CCK-8法、集落形成实验、5-乙炔基-2'-脱氧尿苷(EdU)染色法检测C20对人肝癌HepG2细胞增殖的影响;通过彗星实验检测C20(10μmol·L-1)对HepG2细胞DNA损伤的影响;通过流式细胞术检测C20(5、10μmol·L-1)对HepG2细胞周期阻滞的影响;通过Hoechst染色和流式细胞术检测C20(5、10 μmol·L-1)对HepG2细胞凋亡的影响.借助Western blotting法检测C20(5、10 μmol·L-1)处理对HepG2细胞中与凋亡、DNA损伤、细胞周期阻滞相关蛋白表达水平的调控作用.结果 与对照组比较,C20显著抑制HepG2细胞的活力(P＜0.001),给药48 h的半数抑制浓度(IC50)为7.937 μmol·L-1;5 µmol·L-1 C20能够显著抑制HepG2细胞的集落形成能力(P＜0.01);EdU染色结果显示5、10 μmol·L-1的C20能够抑制人肝癌HepG2细胞的增殖能力;5、10 μmol·L-1的C20显著诱导HepG2细胞G2/M期阻滞(P＜0.001);5、10 μmol·L-1的C20显著促进HepG2细胞凋亡(P＜0.001),并显著上调Caspas-3、Caspase-9以及PARP的剪切水平(P＜0.01);10 μmol·L-1的C20能够诱导HepG2细胞发生DNA损伤,并且5、10 μmol·L-1的C20显著上调γH2AX、p21的蛋白水平(P＜0.01).结论 C20能够造成HepG2细胞发生DNA损伤,上调p21蛋白水平,导致细胞G2/M期阻滞,并进一步诱发凋亡,发挥体外抗肝癌作用.
Abstract
Objective To investigate the in vitro antitumor effects of 2',4'-dihydroxy-3'-methyl-3-methoxychalcone(C20)on human hepatocellular carcinoma HepG2 cells and its potential mechanisms.Methods The effects of C20 on the proliferation of HepG2 cells were detected by CCK8 assay,colony formation assay,and 5-ethynyl-2'-deoxyuridine(EdU)staining assay.The effects of C20(10 μmol·L-1)on DNA damage of HepG2 cells were detected by comet assay.The effects of C20(5,10 μmol·L-1)on cell cycle of HepG2 cells were detected by flow cytometry.The effects of C20(5 and 10 μmol·L-1)on apoptosis of HepG2 cells were detected by Hoechst staining and flow cytometry.The effects of C20(5 and 10 μmol·L-1)on expression levels of proteins related to DNA damage,cell cycle,and apoptosis in HepG2 cells were detected by Western blotting.Results Compared with control group,C20 significantly inhibited the viability of HepG2 cells(P＜0.001),with a half-maximal inhibitory concentration(IC50)of 7.937 μmol·L-1 after 48 h of treatment;5 μmol·L-1 C20 significantly inhibited the colony-forming ability of HepG2 cells(P＜0.01);the EdU staining results showed that 5 and 10 μmol·L-1 of C20 could inhibit the proliferation of human hepatocellular carcinoma HepG2 cells;5 and 10 μmol·L-1 of C20 significantly induced G2/M phase arrest in HepG2 cells(P＜0.001);5 and 10 μmol·L-1 of C20 significantly promoted apoptosis in HepG2 cells(P＜0.001),and significantly upregulated the cleavage levels of Caspas-3,Caspase-9,and PARP(P＜0.01);10 μmol·L-1 of C20 could induce DNA damage in HepG2 cells,and 5 and 10 μmol·L-1 of C20 significantly upregulated the protein levels of γH2AX and p21(P＜0.01).Conclusion We speculate that C20 treatment can induce DNA damage and upregulate p21 expression in HepG2 cells,leading to G2/M arrest and further inducing apoptosis,which is partially responsible for anti-hepatocellular carcinoma activity of C20.
关键词
2',4'-二羟基-3'-甲基-3-甲氧基查耳酮/肝癌/DNA损伤/G2/M周期阻滞/p21Key words
2',4'-dihydroxy-3'-methyl-3-methoxychalcone/hepatocellular carcinoma/DNA damage/G2/M arrest/p21引用本文复制引用
基金项目
国家自然科学基金(82074072)
中央高校基本科研业务费专项(2023-JYB-JBQN-051)
北京中医药大学国家级人才精准培育计划(JZPY202206)
国家中医药局青年岐黄学者支持项目()
出版年
2024
NETL
NSTL
万方数据