摘要
目的建立一种利用细胞电阻抗传感技术(ECIS)进行体外药物筛选与评价的方法。方法利用MTT法测定NCI-H460细胞对紫杉醇注射液、多西他赛注射液、长春瑞滨注射液、注射用培美曲塞二钠及顺铂注射液的敏感性,再分别采用ECIS和MTT法测定紫杉醇、多西他赛、长春瑞滨和培美曲塞二钠4种抗肿瘤药对NCI-H460细胞生长的影响。借助配对t检验、组内相关系数以及Bland-Altman法等对两种方法所测得抑制率进行一致性综合评价,结合电子显微镜下细胞的形态学观察,评估所建立的方法。结果通过MTT法测得NCI-H460细胞对紫杉醇及多西他赛注射液敏感,对长春瑞滨注射液及注射用培美曲塞二钠呈中度敏感,顺铂对NCI-H460细胞株的抑制率无明显变化。紫杉醇、多西他赛、长春瑞滨、培美曲塞二钠4种不同药物对NCI-H460细胞作用的动态过程可通过ECIS动态监测法进行定性定量表征,同时还可反映出不同药物作用的特征性差异。另外,通过分析抑制率数据结果,显示出两种方法一致性较好,结合电子显微镜观察不同时期NCI-H460细胞形态更进一步说明此法的可行性。结论所建立的基于细胞电阻抗分析方法可用于体外药物筛选与评价,可有效地表征药物体外抗肿瘤活性,在肿瘤耐药性及个体化给药研究方面具有广阔的临床应用前景。
Objective To establish a new method for in vitro drug screening and evaluation with electrical cell-substrate impedance sensing （ ECIS ） technology .Methods The sensitivity of human large-cell lung cancer cell line NCI-H460 under the effect of five antitumor drugs including paclitaxel , docetaxel , vinorelbine pemetrexed and cis-platinum was measured by methyl thiazolyl tetrazolium （ MTT） assay.Then, ECIS and MTT assay were respectively used to determine the effect of four antitumor drugs on the growth of NCI-H460 cells.Such data analysis methods as paired t-test, intraclass correlation coefficient and Bland-Altman method were adopted to comprehensively evaluate the consistency of the inhibitiory rates measured by the two methods .Cell morphology was observed under an electron microscope to assess the new method . Results The NCI-H460 cells were sensitive to paclitaxel and docetaxel .The sensitivity of NCI-H460 cells to vinorelbine and pemetrexed was moderate .Under the action of cisplatin , the inhibitory rate of NCI-H460 cells had no obvious change with the increase in drug concentration .ECIS could qualitatively and quantitatively characterize the dynamic information on the effects of the four drugs on NCI-H460 cells.The dynamic biological response spectrum could sensitively reflect the dynamic characteristic information on the inhibitory effects of various drugs .Furthermore, the analysis results of inhibitiory rates showed that the two methods were consistent .The morphologies of NCI-H460 cells in various phases under the electron microscope further indicated the feasibility of the method .Conclusion The ECIS-based method can be used for in vitro drug screening and evaluation, which can effectively characterize the antitumor activity of drugs in vitro.It has good prospects of clinical application in researches of tumor drug resistance and individualized medication .
出处
《军事医学》
CAS
CSCD
北大核心
2016年第11期909-914,共6页
Military Medical Sciences
基金
军事医学科学院军事医学创新基金资助项目(2012CXJJ017)
关键词
非小细胞肺癌
细胞电阻抗传感技术
四甲基偶氮唑盐比色法
non-small cell lung cancer
electrical cell-substrate impedance sensing
methyl thiazolyl tetrazolium
