Ubiquitous mitochondrial creatine kinase downregulated in oral squamous cell carcinoma.
Ubiquitous mitochondrial creatine kinase downregulated in oral squamous cell carcinoma.
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DOI：
10.1038/sj.bjc.6602986
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发表时间：
2006-03-13
影响因子：
8.8
通讯作者：
中科院分区：
文献类型：
--
作者：
在这项研究中，我们进行了二维电泳（2-DE）和基质辅助激光脱附/电离时间飞行质谱，通过比较oscc来源的细胞系与正常口腔角质形成细胞（NOKs）的模式来鉴定与口腔鳞状细胞癌（oscc）发展相关的蛋白质，并发现普遍存在的线粒体肌酸激酶（CKMT1）的下调可能是一个很好的选择。与原代正常口腔角质形成细胞相比，在所有oscc来源的细胞系（n=9）中检测到CKMT1 mRNA和蛋白水平降低。尽管在这些细胞系中，除了外显子8的无义突变外，CKMT1基因的编码区没有发现序列变异，但我们发现CpG岛区域存在频繁的高甲基化。通过实验性去甲基化恢复CKMT1的表达。此外，当我们将CKMT1转染到细胞系中时，它们表现出凋亡表型，但没有侵袭性。在临床样本中，通过免疫组化（52例中有19例（37%））和实时定量RT-PCR（50例中有21例（42%））检测到高频率的CKMT1下调。此外，CKMT1表达状态与肿瘤分化有显著相关性（P<0.0001）。这些结果表明，CKMT1基因在口腔癌发生过程中经常失活，表观遗传机制可能调节其表达缺失，从而导致细胞凋亡受阻。
In this study, we performed two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionisation time of fly mass spectrometry to identify the protein(s) associated with the development of oral squamous cell carcinomas (OSCCs) by comparing patterns of OSCC-derived cell lines with normal oral keratinocytes (NOKs), and found that downregulation of ubiquitous mitochondrial creatine kinase (CKMT1) could be a good candidate. Decreased levels of CKMT1 mRNA and protein were detected in all OSCC-derived cell lines examined (n=9) when compared to those in primary normal oral keratinocytes. Although no sequence variation in the coding region of the CKMT1 gene with the exception of a nonsense mutation in exon 8 was identified in these cell lines, we found a frequent hypermethylation in the CpG island region. CKMT1 expression was restored by experimental demethylation. In addition, when we transfected CKMT1 into the cell lines, they showed an apoptotic phenotype but no invasiveness. In clinical samples, high frequencies of CKMT1 downregulation were detected by immunohistochemistry (19 of 52 (37%)) and quantitative real-time RT–PCR (21 of 50 (42%)). Furthermore, the CKMT1 expression status was significantly correlated with tumour differentiation (P<0.0001). These results suggest that the CKMT1 gene is frequently inactivated during oral carcinogenesis and that an epigenetic mechanism may regulate loss of expression, which may lead to block apoptosis.
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