基于细胞动态生物反应谱的抗肿瘤先导化合物筛选方法研究  

A method of screening the antitumor lead compounds based on the dynamic bio-response profile of cells

作　　者：马丽娜[1,2] 张乐乐[2] 熊吟[2] 韩玉梅[3] 章从恩[2] 郜丹 马莉[4] 鄢丹[2] 肖小河[2] 

机构地区：[1]成都中医药大学药学院,四川成都610075 [2]解放军第302医院全军中药研究所,北京100039 [3]北京市体检中心,北京100077 [4]首都医科大学中医药学院,北京100069

出　　处：《药学学报》2014年第5期695-700,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81274078;81303222;81322052);北京市科技新星人才计划项目(2010B108)

摘　　要：为弥补抗肿瘤活性评估常规断点检测法操作繁琐、灵敏度低等局限,尝试建立基于细胞动态生物反应谱的抗肿瘤先导化合物筛选新方法。依据活细胞具有电阻抗生物传感属性,以细胞指数、半数抑制浓度IC50和脱附曲线为评价指标,采用实时在线分析技术动态监测模式药表柔比星、顺铂、卡铂对HepG2细胞生长的影响,并辅以CCK-8(cell counting kit-8)和显微监视佐证。结果显示,以细胞指数-作用时间关系表征的细胞动态生物反应谱可灵敏地反映模式药抑制HepG2细胞的动态特征效应信息,3种药物的IC50分别为(0.53±0.04)、(9.79±0.26)和(597.00±3.79)μg·mL-1,进一步发现3种药物的脱附曲线差异明显,提示其作用特征可能不同,这与常规检测方法及文献报道一致。研究表明,具有无标记、非侵入、实时动态等特点的细胞动态生物反应谱技术,可定性定量地表征3种药物对HepG2细胞作用的过程动态信息,反映不同药物作用特征性差异,有望为抗肿瘤先导化合物的发现及作用特点或机制阐释提供线索。The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing （RT-CES） system was used to monitor the effect of epirubicin （EPI）, cisplatinum （DDP） and carboplatin （CBP） on the growth of HepG2 cells, with the cell index （CI）, half maximal inhibitory concentration （IC50） and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 （CCK-8） and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.534-0.04, 9.794-0.26 and 597.004-3.79μg.mL-1, respectively. What＇s more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.

关 键 词：先导化合物 活性评价 实时细胞电子传感系统 

分 类 号：R965[医药卫生—药理学]