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Calcein—AM和Annexin V在流式细胞术检测早期细胞凋亡的敏感性比较
  • ISSN号:1001-764X
  • 期刊名称:《临床检验杂志》
  • 时间:0
  • 分类:R329.2[医药卫生—人体解剖和组织胚胎学;医药卫生—基础医学]
  • 作者机构:[1]华中科技大学同济医学院分子医学中心,武汉430030
  • 相关基金:国家自然科学基金(39730270);973肿瘤计划资助项目(G1998051213);卫生部临床重点学科资助项目(20012537).
中文摘要:

目的 比较Calcein-AM与Annexin V用于流式细胞术检测活细胞早期凋亡时的敏感性,建立新的检测细胞早期凋亡的方法。方法将人类急性T淋巴细胞白血病细胞株(Molt-4)经喜树碱(CPT)、紫外线(UV)诱导后,分别采用Calcein-AM及异硫氰酸荧光素标记的膜联蛋白V(Annexin V)染色,然后用流式细胞仪分析凋亡率,通过比较诱导后不同时间点的凋亡率来分析Calcein-AM和Annexin V对于检测细胞早期凋亡的敏感性。结果在喜树碱诱导模型中,加药后2 h即可用Calcein-AM检测到明显的细胞凋亡,而3 h后才能用Annexin V检测到;在紫外线诱导模型中,照射后1 h即可用Calcein-AM检测到明显的细胞凋亡,而要在2 h后才能用Annexin V检测到。结论Calcein-AM用于检测活细胞早期凋亡比Annexin V更为敏感,是一种稳定、可靠、灵敏度高并且较为经济的检测细胞早期凋亡的活细胞染料。

英文摘要:

Objective The current study was designed to compare the sensitivities of Calcein-AM and Annexin V-FITC for early detectionof apoptosis in living cells by flow cytometry and to establish a new method for early detection of apoptosis. Methods The Molt-4 cellline treated with camptothecin (CPT) or ultraviolet(UV) were labeled with Calcein-AM and FITC-conjugated Annexin V respectively,then the apoptosis rate were analyzed by flow cytometry and the performance of Calcein-AM and Annexin V for early detection of apoptosis were compared. Results Our results showed that both probes allowed the detection of apoptotic cells. I~Iowever, Calcein-AM was moresensitive than Annexin V. We could detect the apoptosis induced by CPT in two hours after inducing with Calcein-AM and in threehours after inducing with Annexin V. The apoptosis induced by UV could be detected in one hours after inducing with Calcein-AM buttwo hours after inducing with Annexin V. Conclusion Annexin V is less sensitive than Calcein-AM for detection of early apoptosis, andthe new method for detection of early apoptosis with Calcein-Am is stable,reliable, sensitive and low-cost.

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期刊信息
  • 《临床检验杂志》
  • 北大核心期刊(2011版)
  • 主管单位:江苏省卫生和计划生育委员会
  • 主办单位:江苏省医学会
  • 主编:许文荣
  • 地址:南京市中央路42号
  • 邮编:210008
  • 邮箱:editor@lcjyzz.com
  • 电话:025-83620683
  • 国际标准刊号:ISSN:1001-764X
  • 国内统一刊号:ISSN:32-1204/R
  • 邮发代号:28-104
  • 获奖情况:
  • 国家科技部中国科技论文统计源期刊,中国科学引文数据库医学类核心期刊,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2000版)
  • 被引量:21982