二苯并呋喃-2-基喹啉-8-磺酸盐靶向拓扑异构酶Ⅰ诱导肝癌细胞HepG-2凋亡  

Dibenzofuran-2-ylquinoline-8-sulfonate induces apoptosis of human hepatocellular carcinoma HepG-2 through targeting topoisomerase I

作　　者：张琼 成钟[1] 任吉霞 冯妙 卜浩林 郭建红[1] ZHANG Qiong;CHENG Zhong;REN Ji-xia;FENG Miao;BU Hao-lin;GUJian-hong(Basic Medical College,Shanxi Medical University,Taiyuan 030001,China;College of Life Sciences,Shandong Liaocheng University,Liaocheng 252059,China)

机构地区：[1]山西医科大学基础医学院,太原030001 [2]山东聊城大学生命科学学院,聊城252059

出　　处：《中国新药杂志》2019年第23期2878-2884,共7页Chinese Journal of New Drugs

基　　金：国家自然科学基金青年基金(81502963);山西省科技厅基础科研优秀青年基金(201601D021008)

摘　　要：目的:探讨二苯并呋喃-2-基喹啉-8-磺酸盐(dibenzofuran-2-ylquinoline-8-sulfo-nate,DFQS)以靶向拓扑异构酶Ⅰ(topoisomerase I,TopoⅠ)的方式诱导人肝癌细胞HepG-2凋亡。方法:采用p BR322 DNA质粒超螺旋松弛实验确认DFQS是一种新型的TopoⅠ抑制剂;CCK-8法检测DFQS对HepG-2细胞生存率影响;Hoechst 33258和AO/EB荧光染色法以及流式细胞术探究细胞凋亡变化情况;Western Blot法检测DFQS对HepG-2细胞caspase-3和TopoⅠ蛋白表达影响;利用分子对接阐明DFQS与TopoⅠ相互作用方式。结果:DFQS可抑制TopoⅠ对DNA的催化解旋作用,可显著降低HepG-2细胞存活率,抑制效应呈时间浓度依赖性;DFQS致HepG-2胞核凝集固缩,呈典型凋亡形态学、凋亡率明显提升、caspase-3表达显著增加、TopoⅠ的表达下降;分子模拟结果表明DFQS可结合到TopoⅠ活性区域。结论:DFQS可靶向抑制TopoⅠ活性诱导HepG-2细胞凋亡,发挥其抗癌效应。Objective:To study the effect of dibenzofuran-2-ylquinoline-8-sulfonate(DFQS)on inducing apoptosis of human hepatocellular carcinoma Hep G-2 in a manner of targeting topoisomerase I(TopoⅠ).Methods:DFQS was confirmed as a novel TopoⅠinhibitor using pBR322 DNA plasmid supercoiled relaxation assay.The effect of DFQS on survival rate of HepG-2 cells was analyzed by CCK-8 method.Hoechst 33258,AO/EB fluorescencestaining and flow cytometry were used to detect the change of apoptosis.Western Blot was used to detect the effect of DFQS on the expressions of caspase-3 and Topo I in HepG-2 cells.The interaction between DFQS and Topo I was elucidated using molecular docking method.Results:DFQS inhibited the catalytic unwinding effect of Topo I on DNA and significantly reduced the survival rate of Hep G-2 cells.This inhibitory effect was time-concentrationdependent.DFQS induced HepG-2 nucleus agglutination and pyknosis,showing typical apoptotic morphology.The apoptotic rate and the caspase-3 expression significantly increased,while the expression of Topo I decreased.The results of molecular simulation indicated that DFQS binded to the active region of Topo I.Conclusion:DFQS can induce the apoptosis of HepG-2 cells by inhibiting the activity of Topo I thus exert its anticancer effect.

关 键 词：二苯并呋喃-2-基喹啉-8-磺酸盐 拓扑异构酶Ⅰ 凋亡 HEPG-2 

分 类 号：R967[医药卫生—药理学]