新型β-榄香烯衍生物MDCI对肝癌细胞增殖的抑制作用及机制研究  

Inhibitory effect and mechanism of MDCI,a novelβ-elemene derivative,on the proliferation of hepatocellular carcinoma cells

作　　者：滕佳奕 张一鸣 夏广磊 高慕聪 刘国都 夏勇 TENG Jiayi;ZHANG Yiming;XIA Guanglei;GAO Mucong;LIU Guodu;XIA Yong(School of Clinical Medicine,Jining Medical University,Jining 272013,China;School of Chemistry and Chemical Engineering,Inner Mongolia University,Hohhot 010030,China;School of Medical Engineering,Jining Medical University,Jining 272199,China)

机构地区：[1]济宁医学院临床医学院,济宁272013 [2]内蒙古大学化学化工学院,呼和浩特010030 [3]济宁医学院医药工程学院,济宁272199

出　　处：《济宁医学院学报》2024年第5期365-372,共8页Journal of Jining Medical University

基　　金：国家中医药管理局科技司共建科技项目(G2Y-kJS-SD-2023-097);山东省泰山学者青年专家基金(tsqn201909147);山东省自然科学基金(ZR2023LZL005)。

摘　　要：目的β-榄香烯是我国自主知识产权的创新药,但是其起效浓度偏高,抑癌效率偏低。本研究旨在筛选具有高效抗肝癌活性的β-榄香烯衍生物,并初步揭示其抑制肝癌细胞恶性表型的细胞学机制。方法选取人肝癌细胞珠MHCC97-H作为实验对象,分别用β-榄香烯、β-榄香烯衍生物(MDCC、MCMC、MDCI)处理细胞,通过CCK-8实验对细胞活性变化进行评估。经对比后选择对肝癌细胞活性抑制最强的MDCI进行深入研究:在显微镜下跟踪监测细胞的形态变化,通过细胞划痕实验检测其对细胞迁移能力的影响,采用EdU荧光染色实验判断其对肝癌细胞DNA复制能力的影响,借助流式细胞仪测定其对细胞周期的作用,使用活/死细胞荧光染色法对MDCI诱发细胞死亡作用进行评价,并借助Annexin-V-FITC凋亡试剂盒检测细胞凋亡。结果3种β-榄香烯衍生物对肝癌细胞的抑制能力较β-榄香烯均有不同程度提升,其中MDCI对肝癌细胞的抑制能力最强。随着MDCI浓度和孵育时间的增加,MHCC97-H细胞的活性受到显著抑制,但是迁移能力未发生显著变化(F=2.636,P>0.05)。MDCI可以降低DNA复制速率(F=133.100,P<0.001)进而阻滞细胞周期进入S期(F=224.500,P<0.001)。当提高MDCI浓度达到10μM时,MHCC97-H细胞开始发生细胞死亡,其主要方式为细胞凋亡。结论我们筛选到一种全新的β-榄香烯衍生物—MDCI,对肝癌细胞有出色的抑制效果,在低浓度时主要通过抑制DNA合成阻滞细胞周期,进而抑制细胞过度增殖;在高浓度时,主要通过诱导细胞凋亡(Apoptosis)起到杀伤肿瘤细胞的作用。MDCI具备潜力成为一种新型的肝癌治疗先导化合物,对于丰富肝癌治疗药物线具有积极意义。Objectiveβ-elemene is an innovative drug with independent intellectual property rights in China,but its effective concentration is high and its anticancer efficiency is low.The aim of this study was to screenβ-elemene derivatives with high anti-hepatocellular carcinoma activity and reveal the cytological mechanism of their inhibition of malignant phenotype of hepatocellular carcinoma cells.Methods Human liver cancer cell MHCC97-H was selected as the experimental object,and the cells were treated withβ-elemene,MDCC,MCMC,and MDCI.The cells activities were evaluated by CCK-8 assay.After comparison,we found MDCI had the strongest inhibition on liver cancer cells,and selected MDCI for further study.The morphological changes of the cells were tracked and monitored under the microscope,and the effects on cell migration were detected by cell scratch test.EdU fluorescence staining was used to determine the effects on DNA replication of liver cancer cells.The effects on cell cycle were measured by flow cytometry,and the effects of the cell death-inducing effect of MDCI were evaluated by live/dead cell fluorescence staining.Apoptosis was detected by Annexin-V-FITC apoptosis kit.Results The inhibitory capacity of threeβ-elemene derivatives on hepatocellular carcinoma cells was improved in different degrees compared withβ-elemene,and MDCI had the strongest inhibitory capacity on hepatocellular carcinoma cells.Increasing the concentration and incubation time of MDCI resulted in a significant inhibition of MHCC97-H cell activity,while migration ability remained relatively unchanged(F=2.636,P>0.05).It has been demonstrated that MDCI reduces the rate of DNA replication(F=133.100,P<0.001),thereby blocking cell cycle progression into the S phase(F=224.500,P<0.001).At a concentration of 10μM,MHCC97-H cells began to undergo apoptosis as the primary mode of cell death.Conclusions We have screened a novelβ-elemene derivative,MDCI,which exhibits excellent inhibitory effects on hepatocellular carcinoma cells.At low concentrations,

关 键 词：肝癌 榄香烯 细胞增殖 细胞周期 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤]