Proteome analysis of UV-B-induced anti-apoptotic regulatory factors
Proteome analysis of UV-B-induced anti-apoptotic regulatory factors
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DOI：
10.1562/2004-09-01-ra-297r1.1
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发表时间：
2005-07-01
影响因子：
3.3
通讯作者：
Goto, R
中科院分区：
文献类型：
--
作者：
Ibuki, Y;Naitou, H;Goto, R
众所周知，紫外线（UV）照射可诱导细胞凋亡，其标志性事件是表皮中晒伤细胞的发生。其中DNA被UV照射损伤而未被修复的角化细胞经历作为晒伤细胞的凋亡。然而，我们以前报道过低剂量的UV-B辐射（类似于0.1 J/cm（2））抑制细胞脱离和血清耗竭诱导的凋亡。细胞凋亡的失调在肿瘤进展和恶性化以及促进对癌症治疗的抗性中是重要的。为了更好地了解紫外线辐射的抗凋亡作用，并设计有效的诱导凋亡的方法，我们尝试用双向差异凝胶电泳（DIGE）技术对低剂量UV-B辐射的NIH 3 T3细胞进行凋亡调控分子的蛋白质组分析。在检测到的总共3811个蛋白质点中，发现42个在经历凋亡的细胞和照射后的细胞之间不同。在所选的点中，使用MALDI-TOF/TOF-MS鉴定了25个，其中一些为结构蛋白。虽然没有检测到典型的凋亡相关分子，可能是因为在本研究中使用的凝胶条件下难以识别低分子量的蛋白质，但认为一些蛋白质参与凋亡。本实验中使用的DIGE系统具有发现与细胞凋亡调控相关的新功能蛋白的优势（包括高水平的统计置信度）。
Ultraviolet (UV) irradiation is well known to induce apoptosis, a hallmark event of which is the occurrence of sunburn cells in the epidermis. Keratinocytes in which DNA damaged by UV irradiation is not repaired undergo apoptosis as sunburn cells. However, we have previously reported that low-dose UV-B irradiation (similar to 0.1 J/cm(2)) suppressed the apoptosis induced by cell detachment and serum depletion. Dysregulation of apoptosis is important in tumor progression and malignancy and in promoting resistance to cancer therapy. To develop a better understanding of the antiapoptotic effect of UV irradiation, and to design the effective induction of apoptosis, we tried the proteome analysis of the molecules regulating apoptosis in low-dose UV-B-irradiated NIH3T3 cells, using two-dimensional difference gel electrophoresis (DIGE). Of a total of 3811 protein spots detected, 42 were found to be different between the cells undergoing apoptosis and cells after the irradiation. Of the spots selected, 25 were identified using MALDI-TOF/TOF-MS, some as structural proteins. Although typical apoptosis-related molecules were not detected, possibly because proteins with low molecular weights were difficult to identify in the gel conditions used in this study, some of the proteins were considered to be involved in apoptosis. The DIGE system used in this experiment has advantages (including a high level of statistical confidence) for discovering new functional proteins related to the regulation of apoptosis.