汉防己甲素衍生物HL-49对Bloom DNA解旋酶生物学特性的影响  

Effects of tetrandrine derivative HL-49 on biological property of Bloom DNA helicase

作　　者：刘金河[1] 张望明 晏文涛 向天 罗维 杨凯琳 徐泽权 刘杰麟 LIU Jin-he;ZHANG Wang-ming;YAN Wen-tao;XIANG Tian;LUO Wei;YANG Kai-lin;XU Ze-quan;LIU Jie-lin(National & Guizhou Joint Engineering Lab for Cell Engineering and Biomedicine Technique,Center for Tissue Engineering and Stem Cell Research,Guizhou Province Key Lab for Regenerative Medicine,Guizhou Medical University,Guiyang 550004,China;Dept of Immunology, College of Basic Medicine,Guizhou Medical University,Guiyang 550025,China;The First Affiliated Hospital of Guizhou University of Chinese Medicine,Guiyang 550001,China;School of Clinical Laboratory Medicine,Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学细胞工程生物医药技术国家地方联合工程实验室,组织工程与干细胞实验中心,贵州省再生医学重点实验室,贵州贵阳550004 [2]贵州医科大学基础医学院免疫教研室,贵州贵阳550025 [3]贵州中医药大学第一附属医院检验科,贵州贵阳550001 [4]贵州医科大学检验学院,贵州贵阳550004

出　　处：《中国药理学通报》2022年第2期209-214,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81360349);贵州省教育厅青年科技人才成长项目(黔教合KY字[2018]177);贵州省科技支撑项目(黔科合支撑[2020]4Y120号);贵州省科学技术基金(黔科合基础[2017]1152);环境污染与疾病监控教育部重点实验室开放课题基金资助项目(黔教合KY字[2019]045)。

摘　　要：目的汉防己甲素衍生物是一类双苄基异喹啉类化合物,具有明显的抗肿瘤活性。研究汉防己甲素衍生物HL-49对Bloom DNA解旋酶(bloom DNA helicase,BLM)构象和生物学活性的影响对于探索其抗肿瘤作用机制具有重要意义。方法采用紫外光谱扫描研究药物HL-49对BLM DNA解旋酶构象的影响,荧光偏振技术和孔雀绿-磷钼酸铵比色法分析HL-49对BLM解旋酶的DNA结合活性、DNA解链活性、ATPase活性。结果紫外光谱扫描结果显示,HL-49对BLM DNA解旋酶的三维构象影响不明显。荧光偏振法结果显示,HL-49对BLM解旋酶的DNA(dsDNA/ssDNA)结合活性的影响不明显,但随浓度的增加,与DNA相互结合的能力越强(P<0.01)。随着HL-49浓度的增加,BLM DNA解旋酶的DNA解链能力下降,K_(obs)值逐渐降低。孔雀绿-磷钼酸铵比色结果说明,随着HL-49浓度的增加或作用时间延长,HL-49能明显抑制BLM DNA解旋酶的ATPase活性。结论汉防己甲素衍生物HL-49通过与DNA的可逆性结合,抑制BLM DNA解旋酶的ATPase活性和DNA解链活性。Aim To study the effect of tetrandrine derivative HL-49 on the conformation and biological activity of Bloom helicase(BLM),and to explore its anti-tumor mechanism.Methods The effect of HL-49 on the conformation of BLM helicase was studied by ultraviolet spectroscopy.The effects of HL-49 on DNA binding activity,DNA chain dissociation activity and ATPase activity of HL-49 on BLM DNA helicase were analyzed by fluorescence polarization and malachite green-ammonium phosphomolybdate colorimetric method.Results HL-49,a tetrandrine derivative,indirectly inhibited the ATPase activity of BLM DNA helicase and DNA unwinding activity by reversible binding with DNA.The results of fluorescence polarization experiments showed that HL-49 could not affect the binding activity of BLM DNA helicase to DNA(dsDNA/ssDNA),but could bind to DNA in a concentration-dependent manner(P<0.01).With the increase of HL-49 concentration,the DNA unwinding ability of BLM DNA helicase decreased,and the K_(obs) value decreased gradually.The results of malachite green-ammonium phosphomolybdate colorimetry showed that HL-49 could significantly inhibit the ATPase activity of BLM DNA helicase.Conclusions HL-49 can inhibit the ATPase activity and DNA unwinding activity of BLM DNA helicase by the reversible binding with DNA.

关 键 词：汉防己甲素衍生物 Bloom解旋酶 生化活性 构象 荧光偏振 ATP酶活性 

分 类 号：R284.1[医药卫生—中药学] R342.3[医药卫生—中医学]