Enhancement of UVB-induced apoptosis by apigenin in human keratinocytes and organotypic keratinocyte cultures
Enhancement of UVB-induced apoptosis by apigenin in human keratinocytes and organotypic keratinocyte cultures
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DOI：
10.1158/0008-5472.can-07-2763
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发表时间：
2008-04-15
期刊：
影响因子：
11.2
通讯作者：
Pelling, Jill C.
中科院分区：
文献类型：
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作者：
Abu-Yousif, Adrian O.;Smith, Kimberly A.;Pelling, Jill C.
将生物类黄酮 4',5,7-三羟基黄酮（芹菜素）局部涂抹在小鼠皮肤上，可有效降低由 UVB 暴露引起的皮肤肿瘤的发生率和大小。作为化学预防化合物的能力表明芹菜素治疗改变了由 UVB 暴露引发的分子事件；然而，芹菜素治疗对 UVB 照射的角质形成细胞的影响尚不完全清楚。在本研究中，我们使用三种人类角质形成细胞模型来研究芹菜素治疗对 UVB 诱导的细胞凋亡的影响：HaCaT 人类角质形成细胞、从人类新生儿包皮分离的原代角质形成细胞培养物和人类器官型角质形成细胞培养物。将每个角质形成细胞模型暴露于中等剂量的UVB（300-1,000 J/m(2)），然后用芹菜素（0-50 mu mol/L）处理，并收获并通过聚（ADP）核糖聚合酶裂解的蛋白质印迹分析、流式细胞仪的膜联蛋白-V染色和/或晒伤细胞的存在来评估细胞凋亡。在每个测试的模型中，芹菜素处理可将 UVB 诱导的细胞凋亡增强 2 倍以上。当角质形成细胞暴露于 UVB 时，芹菜素处理会刺激 Bax 定位的变化，并增加线粒体中细胞色素 c 的释放。与单独照射 UVB 相比。抗凋亡蛋白 Bcl-2 的过度表达和 Fas 相关死亡结构域的显性失活形式的表达导致芹菜素增强 UVB 诱导的细胞凋亡的能力降低。这些结果表明芹菜素处理增强 UVB 诱导的细胞凋亡涉及内在和外在的细胞凋亡途径。芹菜素增强 UVB 诱导的细胞凋亡的能力可以部分解释芹菜素的光化学预防作用。
Topical application of the bioflavonoid 4',5,7-trihydroxyflavone (apigenin) to mouse skin effectively reduces the incidence and size of skin tumors caused by UVB exposure. The ability to act as a chemopreventive compound indicates that apigenin treatment alters the molecular events initiated by UVB exposure; however, the effects of apigenin treatment on UVB-irradiated keratinocytes are not fully understood. In the present study, we have used three models of human keratinocytes to study the effect of apigenin treatment on UVB-induced apoptosis: HaCaT human keratinocyte cells, primary keratinocyte cultures isolated from human neonatal foreskin, and human organotypic keratinocyte cultures. Each keratinocyte model was exposed to a moderate dose of UVB (300-1,000 J/m(2)), then treated with apigenin (0-50 mu mol/L), and harvested to assess apoptosis by Western blot analysis for poly(ADP)ribose polymerase cleavage, annexin-V staining by flow cytometry, and/or the presence of sunburn cells. Apigenin treatment enhanced UVB-induced apoptosis > 2-fold in each of the models tested. When keratinocytes were exposed to UVB, apigenin treatment stimulated changes in Bax localization and increased the release of cytochrome c from the mitochondria. compared with UVB exposure alone. Overexpression of the antiapoptotic protein Bcl-2 and expression of a dominant-negative form of Fas-associated death domain led to a reduction in the ability of apigenin to enhance UVB-induced apoptosis. These results suggest that enhancement of UVB-induced apoptosis by apigenin treatment involves both the intrinsic and extrinsic apoptotic pathways. The ability of apigenin to enhance UVB-induced apoptosis may explain, in part, the photochemopreventive effects of apigenin.