Role of NF-κB/Snail/RKIP Loop in the Response of Tumor Cells to Photodynamic Therapy
Role of NF-κB/Snail/RKIP Loop in the Response of Tumor Cells to Photodynamic Therapy
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DOI：
10.1002/lsm.21095
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发表时间：
2011-09-01
影响因子：
2.4
通讯作者：
Xodo, Luigi E.
中科院分区：
文献类型：
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作者：
Rapozzi, Valentina;Umezawa, Kazuo;Xodo, Luigi E.
背景和目的：光动力疗法（PDT）是一种治疗方式，其疗效取决于几个因素，包括光敏剂的类型、光通量和细胞反应。细胞复发是光动力疗法尚未解决的问题之一。在这项工作中，我们发现，在B78-H1小鼠无色素性黑色素瘤细胞有一个细胞复发和NF-κ B/Snail/RKIP loop.Materials和方法：存活细胞的增殖和迁移进行了分析，通过MTT和伤口划痕试验。流式细胞仪检测脱镁叶绿酸a（Pba）和光（Pba/PDT）处理B78-H1黑色素瘤细胞后ROS/NO水平，Western印迹检测NF-κ B、Snail和RKIP表达。结果：低剂量Pba/PDT处理后，B78-H1细胞能够恢复正常。这与低水平的NO产生相关，其通过NF-κ B途径阻断细胞凋亡。Western blot分析显示，低剂量Pba/PDT增加NF-κ B和抗凋亡Snail的表达，但减少促凋亡RKIP的表达。使用DHMEQ（一种表现为NO供体的NF-κ B抑制剂）研究了NF-κ B在Snail和RKIP调节中所起的作用。DHMEQ使Snail表达减少，RKIP表达增加。当B78-H1细胞用低剂量Pba/PDT和DHMEQ处理时，NO水平强烈增加，结果Snail下调，RKIP上调，如用高剂量Pba/PDT.Conclusion：PDT中的一个主要问题是在接受低剂量PDT的组织区域中发生的细胞拯救。为了最大限度地减少这个问题，并使癌细胞对PDT敏感，我们提出了一种联合治疗方法，其中光敏剂与作用于NF-κ B/Snail/RKIP环的NO供体一起递送。激光外科医学43：575-585，2011年。(c)2011 Wiley-Liss，Inc.
Background and Objective: Photodynamic therapy (PDT) is a therapeutic modality whose efficacy depends on several factors including type of photosensitizer, light fluence and cellular response. Cell recurrence is one of the problems still unsolved in PDT. In this work we found that in B78-H1 murine amelanotic melanoma cells there is a correlation between cell recurrence and the NF-kappa B/Snail/RKIP loop.Materials and Methods: Proliferation and migration of surviving cells were analyzed by MTT and wound-scratch assays. The levels of ROS/NO in B78-H1 melanoma cells treated with pheophorbide a (Pba) and light (Pba/PDT) were measured by FACS, while expression of NF-kappa B, Snail and RKIP were determined by Western blots. The mechanism of cell death was investigated by caspase and microscopy assays.Results: Our data show that after a low-dose Pba/PDT treatment, B78-H1 cells are able to recover. This correlates with a low level of NO production, which blocks apoptosis via NF-kappa B pathway. Western blot analyses showed that a low-dose Pba/PDT increases the expression of NF-kappa B and anti-apoptotic Snail, but reduces the expression of pro-apoptotic RKIP. The role played by NF-kappa B in the modulation of Snail and RKIP was investigated using DHMEQ: a NF-kappa B inhibitor which behaves as NO donor. DHMEQ caused a decrease of Snail and an increase of RKIP expression. When B78-H1 cells were treated with a low dose Pba/PDT and DHMEQ, the NO level strongly increased, with the result that Snail was down-regulated and RKIP was upregulated, as observed with a high-dose Pba/PDT.Conclusion: One major problem in PDT is the cellular rescue occurring in tissue regions receiving a low-dose PDT. To minimize this problem and sensitize cancer cells to PDT we propose a combined treatment in which the photosensitizer is delivered with a donor of NO acting on the NF-kappa B/Snail/RKIP loop. Lasers Surg. Med. 43: 575-585, 2011. (c) 2011 Wiley-Liss, Inc.