尾叶香茶菜丙素对肝癌耐药株HepG2/ADM细胞体外活性的影响  

In vitro activity of Kamebakaurin on hepatocellular carcinoma resistant strain HepG2/ADM cells

作　　者：王欣玉 裴晓东 何志龙 林佳雯 黎永卓 陈文卿 刘小玲[1,2] 蒋利和 WANG Xinyu;PEI Xiaodong;HE Zhilong;LIN Jiawen;LI Yongzhuo;CHEN Wenqing;LIU Xiaoling;JIANG Lihe(College of Light Industry and Food Engineering,Guangxi University,Nanning,Guangxi Zhuang Autonomous Region 530004,China;State Key Laboratory of Bioactive Substance and Function of Natural Medicines,Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China;Medical College,Guangxi University,Nanning,Guangxi Zhuang Autonomous Region 530004,China)

机构地区：[1]广西大学轻工与食品工程学院,广西壮族自治区南宁530004 [2]中国医学科学院/北京协和医学院药物研究所,天然药物活性物质与功能国家重点实验室,北京100050 [3]广西大学医学院,广西壮族自治区南宁530004

出　　处：《安徽医药》2021年第11期2131-2135,共5页Anhui Medical and Pharmaceutical Journal

基　　金：天然药物活性物质与功能国家重点实验室开放课题(GTZK201810)。

摘　　要：目的探究尾叶香茶菜丙素(Kamebakaurin)对HepG2/ADM(肝癌细胞耐阿霉素耐药株)细胞体外活性。方法通过2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺酸苯)-2H-四唑单钠盐(CCK-8法)测定尾叶香茶菜丙素和阿霉素(ADM)对HepG2/ADM细胞的生长活力抑制率;通过流式细胞术测定尾叶香茶菜丙素对细胞周期和细胞凋亡的影响;运用Transwell小室实验和划痕愈合实验检验尾叶香茶菜丙素对HepG2/ADM细胞迁移能力影响,用实时荧光定量PCR检测耐药相关基因表达,通过蛋白印迹法(Western blotting)分析PTEN-AKT信号通路。结果尾叶香茶菜丙素对HepG2/ADM细胞的IC50值为62.96µmol/L,ADM在1 mg/mL(184µmol/L)只有26.5%抑制率,说明HepG2/ADM细胞耐药株对尾叶香茶菜丙素敏感度高于阿霉素;尾叶香茶菜丙素诱导肝癌细胞HepG2/ADM细胞的凋亡,阻滞细胞在G2期,且呈浓度依赖性;尾叶香茶菜丙素具有抑制HepG2/ADM细胞穿过小室的能力和平板愈合能力,显示尾叶香茶菜丙素可抑制其迁移能力;Western blotting显示尾叶香茶菜丙素通过抑制MDR1和PENT-AKT通路发挥以上功能。结论尾叶香茶菜丙素抑制MDR1蛋白表达从而逆转HepG2/ADM细胞对药物的敏感性,通过抑制PENT-AKT通路的激活从而促进HepG2/ADM细胞的凋亡并抑制其迁移能力。Objective To investigate the activity of Kamebakaurin on HepG2/ADM cells in vitro.Methods CCK-8 kit was used to determine the cell viability of HepG2/ADM cells treated by Kamebakaurin and adriamycin(ADM).The effects of Kamebakaurin on cell cycle and apoptosis were determined by flow cytometry.Transwell assay and wound healing experiments were used to measure the migratory and invasive ability on HepG2/ADM cells,drug resistance related genes were detected by real-time quantitative PCR.PTENAKT pathway was assayed by western blotting.Results The IC50 value was 62.96μmol/L of Kamebakaurin on HepG2/ADM cells,while 1 mg/mL ADM(184μmol/L)on HepG2/ADM cells had an inhibition of 26.5%,therefore HepG2/ADM cells were more sensitive to Kamebakaurin than ADM.We also found Kamebakaurin promoted the apoptosis of HepG2/ADM cells,arrested cell cycle in G2 phase and showed concentration dependence.Moreover,Kamebakaurin inhibited the migratory ability of HepG2/ADM cells by sup‐pressing the expression of MDR1 and PTEN-AKT pathway.Kamebakaurin inhibited the ability of HepG2/ADM cells to pass the tran‐swell and to heal wound gap,indicating that Kamebakaurin could suppress its migration ability.Conclusion Kamebakaurin reverses the sensitivity to drug of HepG2/ADM cells by inhibiting MDR1 protein expression,promoting the apoptosis of HepG2/ADM cells and suppressing its migration ability by inactivating PTEN-AKT pathway.

关 键 词：香茶菜属 肝肿瘤 抗药性 肿瘤 尾叶香茶菜丙素 PENT-AKT通路 凋亡 迁移 

分 类 号：R285[医药卫生—中药学]