芹菜素对MHCC97H源性球细胞对5-氟尿嘧啶敏感性的影响  

Apigenin up-regulates expression of miR-34a-5p and enhances sensitivity to fluorouracil in MHCC97H-derived spheres

作　　者：乐伊婕 曹建国[1] 杨小红[1] 冯星[1] LE Yi-jie;CAO Jian-guo;YANG Xiao-hong;FENG Xing(Department of Pharmaceutical Sciences,Medical College of Hunan Normal University,Changsha 410013)

机构地区：[1]湖南师范大学医学院药学系,长沙410013

出　　处：《中南药学》2024年第6期1566-1571,共6页Central South Pharmacy

基　　金：国家自然科学基金项目(No.82074075)。

摘　　要：目的 研究芹菜素(API)对人肝细胞癌MHCC97H源性球细胞(MH-SFC)对5-氟尿嘧啶(5-FU)的敏感性的影响,并探讨其分子机制。方法 应用无血清干细胞培养基超低黏附培养获得MH-SFC。CCK-8检测5-FU或API抑制MH-SFC和MHCC97H细胞存活力的半数抑制浓度(IC_(50))。实时定量聚合酶链反应(qRT-PCR)分析miR-34a-5p表达水平。API(10.0、40.0 μmol·L^(-1))预处理或API(40.0 μmol·L^(-1))联合miR-34a-5p抑制物(Anti-34a)共处理MH-SFC,随后测定5-FU的IC_(50)。Western blot分析MH-SFC的FoxM1及c-Myc蛋白表达水平。结果 与MHCC97H细胞相比,MH-SFC 的5-FU的IC_(50)增高。API优先抑制MH-SFC细胞存活力。API(10.0、40.0 μmol·L^(-1))预处理降低MH-SFC的5-FU的IC_(50)。与MHCC97H细胞相比,MH-SFC更低表达miR-34a-5p,同时,API上调miR-34a-5p表达。Anti-34a能消除API增强MH-SFC的5-FU敏感性和上调miR-34a-5p表达作用。此外,API(10.0、40.0 μmol·L^(-1))下调MH-SFC的FoxM1及c-Myc蛋白表达;Anti-34a能消除API下调FoxM1及c-Myc蛋白表达效应。结论 API增强MH-SFC的5-FU敏感性,其分子机制与上调miR-34a-5p表达,继而阻断FoxM1/c-Myc通路相关。Objective To determine the effect of apigenin(API)on the sensitivity of human hepatocellular carcinoma MHCC97H-derived spheres(MH-SFC)to 5-fluorouracil(5-FU),and related mechanism.Methods MH-SFC were obtained from MHCC97H cell line by sphere formation assay with serum-free stem cell medium in ultra low attachment plates.The IC_(50) of 5-FU or API on the cell viability in MH-SFC and MHCC97H cells was assessed by cell counting kit-8(CCK-8)assay.MiR-34a-5p expression level was measured by qRT-PCR.After pre-treatment with API(10.0 and 40.0μmol·L^(-1))or co-treatment with API(40.0μmol·L^(-1))and miR-34a-3p inhibitor(Anti-34a),IC_(50) of 5-FU was determined in MH-SFC.The expressions of FoxM1 and c-Myc protein were analyzed by Western blot.Results IC_(50) of 5-FU was elevated in MH-SFC compared with MHCC97H cells.API preferentially inhibited the cell viability of MH-SFC.IC_(50) of 5-FU was reduced in MH-SFC by pre-treatment of API(10.0 and 40.0μmol·L^(-1)).Compared with the MHCC97H cells,MHSFC had lower expression of miR-34a-5p as well as API up-regulated the expression of miR-34a-5p.Anti-34a rescued the effect of API on the sensitivity to 5-FU and expression of miR-34a-5p in MH-SFC.In addition,API(10.0 and 40.0μmol·L^(-1))down-regulated the expressions of FoxM1 and c-Myc protein in MH-SFC.Anti-34a decreased the down-regulation of expressions of FoxM1 and c-Myc protein in MH-SFC by API.Conclusion API can enhance the sensitivity of MH-SFC to 5-FU,the mechanism may be associated with up-regulating miR-34a-5p expression,and subsequently interrupting FoxM1/c-Myc signaling pathway.

关 键 词：芹菜素 肝细胞癌 肿瘤干细胞 5-氟尿嘧啶 化疗敏感性 miR-34a-5p FOXM1 C-MYC 

分 类 号：R285[医药卫生—中药学]