Flow cytometric quantification of UV-induced cell death in a human squamous cell carcinoma-derived cell line: dose and kinetic studies
Flow cytometric quantification of UV-induced cell death in a human squamous cell carcinoma-derived cell line: dose and kinetic studies
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DOI：
10.1016/s1011-1344(98)00127-4
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发表时间：
1998-07-10
影响因子：
5.4
通讯作者：
Trautinger, F
中科院分区：
文献类型：
--
作者：
Schindl, A;Klosner, G;Trautinger, F
暴露于紫外线 (UV) 辐射和光化疗会诱导表皮细胞凋亡。在这项研究中，通过流式细胞术测量了膜联蛋白 V 结合和碘化丙啶 (PI) 摄取，以评估紫外线诱导的人鳞状细胞癌衍生细胞系 A 431 中的细胞死亡。UVA、UVA1、UVB、窄带 UVB (311 nm) 的生理和治疗相关剂量以及使用 100 ng/ml 8-甲氧基补骨脂素 (8-MOP) 和 WA 或已应用UVA1（PUVA或PUVA1）。 UVA1 和 WA 的剂量范围为 g 至 96 J/cm(2)，WE 的剂量范围为 8 至 128 mJ/cm(2)，窄带 UVB (311 nm) 的剂量范围为 256 至 4096 mJ/cm(2)，光化疗的剂量范围为 1 至 16 J/cm(2)。结果表明，膜联蛋白 V 结合量（早期细胞凋亡的衡量标准）以及 PI 摄取（最终细胞死亡的参数）与所施加的紫外线剂量严格相关。在所有方案中，在 UV 暴露后 12 小时记录膜联蛋白 V 阳性细胞的峰值，随后 PI 摄取细胞增加，峰值出现在 UVA 和 UVA1 后 24 小时，以及 PUVA、PUVA1、UVB 和窄带 UVB 后 48 小时。为了比较不同波长和光源的效果，根据 PI 摄取确定的 50% 细胞死亡诱导计算剂量当量。 UVA 和 UVA1 的等效能量为 96 J/cm(2)，PUVA 和 PUVA1 的等效能量为 16 J/cm(2)，UVB 的等效能量为 256 mJ/cm(2)，窄带 UVB 的等效能量为 2048 mJ/cm(2)。我们的结果表明膜联蛋白 V/PI 双染色是量化 UV 诱导的细胞死亡的合适方法。此外，它们为进一步研究紫外线诱导的细胞凋亡的机制和修饰提供了基础。 (C) 1998 Elsevier Science S.A. 保留所有权利。
Exposure to ultraviolet (UV) radiation and photochemotherapy induces apoptotic cell death in epidermal cells. In this study annexin V binding and propidium iodide (PI) uptake have been measured by flow cytometry to evaluate W-induced cell death in the human squamous cell carcinoma-derived cell line A 431. Physiological and therapeutical relevant doses of UVA, UVA1, UVB, narrow-band UVB (311 nm) and photochemotherapy using 100 ng/ml of 8-methoxypsoralen (8-MOP) with WA or UVA1 (PUVA or PUVA1) have been applied. Doses ranged from g to 96 J/cm(2) for UVA1 and WA, from 8 to 128 mJ/cm(2) for WE, from 256 to 4096 mJ/cm(2) for narrow-band UVB (311 nm) and from 1 to 16 J/cm(2) for photochemotherapy. Results show that the amount of annexin V binding, a measure of early apoptosis, as well as PI uptake, a parameter of ultimate cell death, are strictly correlated with the applied UV dose. Peak values of annexin V-positive cells are noted 12 h after UV exposure in all protocols and are followed by an increase of PI-uptaking cells with peak values at 24 h after UVA and UVA1, and 48 h after PUVA, PUVA1, UVB and narrow-band UVB. To compare the effect of different wavelengths and light sources, dose equivalents are calculated based on the induction of 50% cell death las determined by PI uptake). The equivalents are 96 J/cm(2) for UVA and UVA1, 16 J/cm(2) for PUVA and PUVA1, 256 mJ/cm(2) for UVB and 2048 mJ/cm(2) for narrow-band UVB. Our results establish annexin V/PI double staining as an appropriate method for the quantification of UV-induced cell death. Moreover, they provide a basis for further investigations concerning mechanisms and modifications of UV-induced apoptosis. (C) 1998 Elsevier Science S.A. All rights reserved.