Altered gene expression in melanocytes exposed to 4-tertiary butyl phenol (4-TBP): upregulation of the A2b adenosine receptor 1.
Altered gene expression in melanocytes exposed to 4-tertiary butyl phenol (4-TBP): upregulation of the A2b adenosine receptor 1.
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暴露于 4-叔丁基苯酚 (4-TBP) 的黑素细胞中基因表达发生改变：A2b 腺苷受体 1 上调。
DOI：
10.1046/j.1523-1747.1999.00756.x
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Boissy,RE
中科院分区：
文献类型：
--
作者：
LePoole,IC;Yang,F;Brown,TL;Cornelius,J;Babcock,GF;Das,PK;Boissy,RE
暴露于酚类药物有助于职业性白癜风的发展。建议作为leukodermain体内的致病因素，对位取代苯酚4-叔丁基苯酚被选择来研究负责从对这些试剂敏感的个体的表皮中选择性地消失黑素细胞的早期细胞事件。为此，在将细胞暴露于250 μM 4-叔丁基苯酚或仅暴露于溶媒12 h后，对从三种单独培养物中分离的黑素细胞mRNA进行差异显示。克隆代表差异表达信息的cDNA片段，随后通过反向斑点印迹法确认。比对分析表明，L30核糖体蛋白的上调的治疗，可能反映了改变水平的蛋白质合成的压力。此外，暴露于4-叔丁基苯酚后上调的基因序列被鉴定为A2 b受体（腺苷的P1受体）。该基因的差异表达在RNA酶保护测定中得到证实。通过逆转录-聚合酶链反应，该基因显示在角质形成细胞和成纤维细胞中表达。流式细胞术证实了在黑素细胞和成纤维细胞中的差异表达，但在角质形成细胞中没有。有趣的是，已有报道P1嘌呤受体刺激可诱导细胞凋亡。这与其他地方报告的结果一致，这些结果表明4-叔丁基苯酚诱导人黑素细胞凋亡，以及本研究中在暴露于250 μM 4-叔丁基苯酚72 h的细胞中观察到的形态学变化。总之，差异显示是有用的，以建立参与细胞反应的酚类药物的黑素细胞成分。
Exposure to phenolic agents contributes to the development of occupational vitiligo. Proposed as a causative factor for leukodermain vivo,the para-substituted phenol 4-tertiary butyl phenol was chosen to investigate early cellular events responsible for selective disappearance of melanocytes from the epidermis of individuals sensitive to such agents. To this end, differential display of melanocyte mRNA isolated from three separate cultures was performed following a 12 h exposure of cells to 250 μM 4-tertiary butyl phenol or to vehicle alone. Fragments of cDNA representing differentially expressed messages were cloned and subsequently confirmed by reverse dot blotting. Alignment analysis revealed that the L30 ribosomal protein was upregulated by the treatment, potentially reflecting altered levels of protein synthesis in response to stress. In addition, a gene sequence upregulated following exposure to 4-tertiary butyl phenol was identified as the A2breceptor (a P1 receptor for adenosine). Differential expression of this gene was confirmed in an RNase protection assay. By reverse transcription–polymerase chain reaction, the gene was shown to be expressed in keratinocytes and fibroblasts as well. Flow cytometry confirmed differential expression in melanocytes and fibroblasts, but not in keratinocytes. Interestingly, it has been reported that P1 purinoceptor stimulation can induce apoptosis. This is in concordance with results reported elsewhere demonstrating induction of apoptosis by 4-tertiary butyl phenol in human melanocytes, as well as with morphologic changes observed in this study in cells exposed to 250 μM 4-tertiary butyl phenol for 72 h. In conclusion, differential display is useful to establish melanocyte components involved in the cellular response to phenolic agents.