355 nm和407 nm激光激发Hoechst 33342检测细胞凋亡的比较研究_《华西医学》

作者：

李雪 ¹ , 吴亚兰 ¹ , 黄鑫 ² , 黄巧容 ¹ ,  孟文彤 ¹

1四川大学华西医院干细胞生物学研究室（成都，610041）;2中国科学院成都生物研究所（成都，610041）;

关键词：

355 nm激光 407 nm激光流式细胞仪凋亡 Hoechst 33342

DOI：

10.7507/1002-0179.20130274

视频：

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目的　比较使用流式细胞仪355 nm和407 nm激光器激发Hochest33342检测细胞凋亡。方法　通过ATO药物诱导急性早幼粒白血病细胞（NB4）及血清饥饿法诱导人肺癌细胞（NCl-H292）细胞凋亡，取24、48 h时间点收集细胞，进行Hoechst33342-碘化丙啶（PI）双染，分别在配置有两种激光器的流式细胞仪上检测细胞凋亡。结果　细胞经处理后24 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（28.20 ± 4.80）%；NCl-H292细胞凋亡率Hoechst33342+/PI-：（22.47 ± 2.78）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（25.10 ± 6.19）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：20.47 ± 1.46%。处理后48 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（33.60 ± 3.75）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（26.77 ± 1.16）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（29.47 ± 2.33）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（31.47 ± 3.05）%。两种细胞处理后比处理前凋亡率明显升高，但355 nm激光器与407 nm激光器检测的凋亡结果差异不明显（P＞0.05）。结论　407 nm激光器激发Hoechst33342可检测细胞凋亡。

引用本文： 李雪,吴亚兰,黄鑫,黄巧容,孟文彤. 355 nm和407 nm激光激发Hoechst 33342检测细胞凋亡的比较研究. 华西医学, 2013, 28(6): 875-878. doi: 10.7507/1002-0179.20130274 复制

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13.	Shen ZX, Chen GQ, Ni JH, et al. Use of Arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients[J]. Blood, 1997, 89(9): 3354-3360.
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1. 　Wlodkowic D, Telford W, Skommer J, et al. Apoptosis and beyond: cytometry in studies of programmed cell death [J]. Methods Cell Biol, 2011, 103: 55-98.
2. 　Latt SA, Stetten G. Spectral studies on 33258 Hoechst and related bisbenzimidazole dyes useful for fluorescent detection of deoxyribonucleic acid synthesis[J]. J Histochem Cytochem, 1976, 24(1): 24-33.
3. 　Veremes I, Haanen C, Reutelingsperger C. Flow cytometry of apoptotic cell death[J]. J Immunol Methods, 2000, 243(1-2): 167-190.
4. 　徐晓雪, 邓君, 邹林樾, 等. 407nm激光流式细胞仪检测小鼠骨髓侧群细胞效果研究[J]. 继续医学教育, 2011, 25(11): 63-66.
5. 　Simpson C, Pearce DJ, Bonnet D, et al. Out of the blue: a comparison of Hoechst side population (SP) analysis of murine bone marrow using 325, 363 and 407 nm excitation sources[J]. J Immunol Methods, 2006, 310(1-2): 171-181.
6. 　Telford WG, Frolova EG. Discrimination of the hoechst side population in mouse bone marrow with violet and near-ultraviolet laser diodes[J]. Cytometry A, 2004, 57(1): 45-52.
7. 　van Engeland M, Nieland LJ, Ramaekers FC, et al. Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure[J]. Cytometry, 1998, 31(1): 1-9.
8. 　Komoriya A, Packard BZ, Brown MJ, et al. Assessment of caspase activities in intact apoptotic thymocytes using cell-permeable fluorogenic caspase substrates[J]. J Exp Med, 2000, 191(11): 1819-1828.
9. 　Mathur A, Hong Y, Kemp BK, et al. Evaluation of fluorescent dyes for the detection of mitochondrial membrane potential changes in cultured cardiomyocytes[J]. Cardiovasc Res, 2000, 46(1): 126-138.
10. 　Mocharla R, Mocharla H, Hodes ME. A novel, sensitive fluorometric staining technique for the detection of DNA in RNA preparations[J]. Nucleic Acids Res, 1987, 15(24): 10589.
11. 　王文静. 侧群细胞的研究进展[J]. 国际口腔医学杂志, 2011, 38(6): 665-669.
12. 　Sanz MA, Grimwade D, Tallman MS, et al. Management of acute promyelocytic leukemia: recommendations from an expert panel on behalf of the European LeukemiaNet[J]. Blood, 2009, 113(9): 1875-1891.
13. 　Shen ZX, Chen GQ, Ni JH, et al. Use of Arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients[J]. Blood, 1997, 89(9): 3354-3360.
14. 　Liu JW, Chandra D, Rudd MD, et al. Induction of prosurvival molecules by apoptotic stimuli: involvement of FOXO3a and ROS[J]. Oncogene, 2005, 24(12): 2020-2031.

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355 nm和407 nm激光激发Hoechst 33342检测细胞凋亡的比较研究

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《华西医学》

355 nm和407 nm激光激发Hoechst 33342检测细胞凋亡的比较研究

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