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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 method of screening and evaluating in vitro drugs by using cell impedance technique (ECIS). Methods The sensitivity of NCI-H460 cells to paclitaxel injection, docetaxel injection, vinorelbine injection, pemetrexed disodium injection and cisplatin injection were determined by MTT assay. ECI and MTT The effects of paclitaxel, docetaxel, vinorelbine and pemetrexed disodium on the growth of NCI-H460 cells were determined. By means of paired t-test, intra-group correlation coefficient and Bland-Altman method, the inhibitory rates of the two methods were evaluated comprehensively, and the established methods were evaluated by morphological observation under the electron microscope. Results MTT assay showed that NCI-H460 cells were sensitive to paclitaxel and docetaxel injection, moderately sensitive to vinorelbine injection and pemetrexed disodium injection, and the effect of cisplatin on NCI-H460 cell line No significant change in inhibition rate. Paclitaxel, docetaxel, vinorelbine, pemetrexed disodium 4 different drugs on the dynamic process of NCI-H460 cells by ECIS dynamic monitoring of qualitative and quantitative characterization, but also reflect the different drug effects Characteristic differences. In addition, by analyzing the results of inhibition rate data, it shows that the consistency of the two methods is good. The observation of the morphology of NCI-H460 cells by electron microscopy further demonstrates the feasibility of this method. Conclusion The established method based on cell impedance analysis can be used to screen and evaluate drug in vitro. It can effectively characterize the antitumor activity of the drug in vitro and has broad clinical application prospects in tumor drug resistance and individualized drug delivery.