摘要
目的:探究载脂蛋白M(ApoM)/线粒体凋亡在肝癌中的作用,以及人参皂苷Rb1(Rb1)通过ApoM/线粒体凋亡途径对肝癌影响的潜在机制。方法:生物信息学分析筛选ApoM在肝癌中的差异性表达及临床验证;分子对接技术确定Rb1与ApoM的靶向结合,体外培养人肝癌细胞(HepG2),并进行细胞活力检测(CCK-8)筛选Rb1最佳干预浓度,克隆实验用于检测HepG2细胞的增殖情况,划痕实验用于检测HepG2细胞的迁移能力,采用蛋白质印迹法(Western Blotting)检测ApoM及线粒体凋亡相关基因Bcl2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl2)、胱天蛋白酶-3(Caspase-3)、胱天蛋白酶-9(Caspase-9)蛋白表达,实时萤光定量聚合酶链式反应(RT-qPCR)法检测ApoM及线粒体凋亡相关基因Bax、Bcl2、Caspase-3、Caspase-9 mRNA表达水平。结果:生存分析发现ApoM低表达肝癌预后更差,分子对接提示Rb1与ApoM之间具有较高亲和力,克隆及划痕实验提示Rb1有效抑制HepG2细胞增殖,Western Blotting及RT-qPCR验证Rb1可以干预线粒体凋亡相关基因蛋白和mRNA表达。结论:揭示了ApoM/线粒体凋亡在肝癌中发挥重要作用,人参皂苷Rb1可能通过ApoM/线粒体凋亡途径影响肝癌的可能潜在机制。
Objective:To explore the role of apolipoprotein M(ApoM)and mitochondrial apoptosis in liver cancer,and the potential mechanism of ginsenoside Rb1 in affecting liver cancer through the ApoM/mitochondrial apoptosis pathway.Methods:Bioinformatics analysis was used to screen for differential expression of ApoM in liver cancer,followed by clinical validation.Molecular docking determined the targeted binding of ginsenoside Rb1 to ApoM.Human liver cancer cells(HepG2)were cultured in vitro,and cell viability was assessed using the CCK-8 assay to screen for the optimal intervention concentration of ginsenoside Rb1.Clonogenic assays were conducted to evaluate HepG2 cell proliferation,while scratch assays were used to assess the migration ability of HepG2 cells.Protein expression of ApoM and apoptosis-related genes,including Bax,Bcl2,Caspase-3,and Caspase-9,was detected using Western blot.Real-time quantitative PCR(RT-qPCR)was employed to measure mRNA expression levels of ApoM and apoptosis-related genes(Bax,Bcl2,Caspase-3,and Caspase-9).Results:Survival analysis indicated that low ApoM expression correlated with poorer prognosis in liver cancer.Molecular docking suggested a high affinity between ginsenoside Rb1 and ApoM.Clonogenic and scratch assays indicated that ginsenoside Rb1 effectively inhibited HepG2 cell proliferation.Western blot and RT-qPCR confirmed that ginsenoside Rb1 modulated the protein and mRNA expression of mitochondrial apoptosis-related genes.Conclusion:This study reveals the important role of the ApoM/mitochondrial apoptosis pathway in liver cancer,suggesting that ginsenoside Rb1 may influence liver cancer through the ApoM/mitochondrial apoptosis pathway.
作者
王嘉鑫
宋囡
王杰
杨莹
朱敬轩
王群
贾连群
WANG Jiaxin;SONG Nan;WANG Jie;YANG Ying;ZHU Jingxuan;WANG Qun;JIA Lianqun(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《世界中医药》
CAS
北大核心
2024年第17期2578-2583,共6页
World Chinese Medicine
基金
国家自然科学基金项目(82074145,82374423)
辽宁省教育厅自然科学基金项目(L202016)
辽宁省科技厅自然科学基金项目(2020-MS-228)。
