生物技术通报 ›› 2019, Vol. 35 ›› Issue (10): 220-226.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0399

• 技术与方法 • 上一篇    下一篇

实时无标记肿瘤细胞凋亡筛选技术体系的建立

李艳伟, 宋兴辉, 王佳佳, 刘丽, 黄莹莹, 郭春   

  1. 浙江大学医学院生物化学与分子生物学平台,杭州 310058
  • 收稿日期:2019-05-09 出版日期:2019-10-26 发布日期:2019-09-30
  • 作者简介:李艳伟,女,硕士,助理研究员,研究方向:细胞生物学检测;E-mail:lywei@zju.edu.cn
  • 基金资助:
    浙江大学实验技术研究项目(SYB201609)

Establishment of the Real-time and Label-free Screening System for Tumor Cell Apoptosis

LI Yan-wei, SONG Xing-hui, WANG Jia-jia, LIU Li, HUANG Ying-ying, GUO Chun   

  1. Facility for Biochemistry and Molecular Medicine Core Facilities,Zhejiang University School of Medicine,Hangzhou 310058
  • Received:2019-05-09 Published:2019-10-26 Online:2019-09-30

摘要: 将xCELLigence-RTCA细胞分析系统的实时无标记技术和流式细胞术结合,建立实时无标记的肿瘤细胞凋亡筛选技术体系。利用顺铂诱导非小肺癌细胞A549凋亡,使用xCELLigence-RTCA系统连续监测96 h获取A549细胞动态生长曲线图谱,分析图谱确定细胞凋亡最佳检测时间点和半有效抑制浓度(IC50),联合流式细胞术的精准定量技术,分析顺铂作用后A549细胞的凋亡率。结果确定这种实时无标记细胞凋亡筛选技术体系用于肿瘤细胞凋亡筛选的可行性和可靠性。

关键词: 凋亡检测, 实时无标记, 流式细胞术, 有效药物浓度

Abstract: A real-time and label-free tumor cell apoptosis screening system via combining real-time xCELLigence-RTCA analysis system and flow cytometry technology. Cisplatin was used to induce the apoptosis of non-small lung cancer cells A549,and xCELLigence-RTCA system through 96 h continuous monitoring was applied to acquire the dynamic growth curve of A549 cells. The optimal testing time of apoptosis and 50% inhibitory concentration was deduced from the growth curve. Combined with precise quantitative flow cytometry technology,apoptotic rate of A549 cells treated with cisplatin was analyzed. These results confirmed the feasibility and reliability of the real-time and free technology for apoptosis screening of tumor cells.

Key words: apoptosis detection, real-time and label-free technology, flow cytometry, effective drug concentration