Wang Jie, Cai Yan, Xu Hao, Zhao Jun, Xu Xin, Han Ya Ling, Xu Zhi Xiong, Chen Bao Sheng, Hu Hai, Wu Min, Wang Ming Rong
National Laboratory of Molecular Oncology, Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking University of Medical School, Beijing 100021, China.
Cell Res. 2004 Feb;14(1):46-53. doi: 10.1038/sj.cr.7290201.
Migration inhibitory factor-related protein 14 (MRP14) is one of calcium-binding proteins, referred as S100A9. The heterodimeric molecule formed by MRP14 with its partner MRP8 (S100A8) is the major fatty acid carrier in neutrophils. The MRP8/14 complex has been also implicated in the intracellular transport of arachidonic acid and its precursors in keratinocytes. We show here the involvement of MRP14 in human esophageal cancer. In an initial study, mRNA differential display-reverse transcription polymerase chain reaction (DD-PCR) was performed with two esophageal carcinomas, one esophageal adenocarcinoma and matched normal adjacent mucosa. DD-PCR with the arbitrary primer OPA3 showed that one cDNA band was highly expressed in normal tissues, but disappeared or substantially decreased in tumor counterparts. It was later identified to be the 3'-end of migration inhibitory factor-related protein 14 (MRP14). Northern blotting, RT-PCR and Western blotting corroborated the down-regulation of MRP14 in 58/64 squamous cell carcinomas and 2/2 adenocarcinomas as compared with adjacent normal epithelia of the esophagus. MRP14 was undetectable in 3/3 esophageal-carcinoma cell lines. Immunochemistry demonstrated that expression of MRP14 was restricted to normal esophageal epithelia. No mutation was found in the genomic DNA of the MRP14 gene by PCR and directed DNA sequencing. Our finding suggested that the reduction of MRP14 expression is a frequent event in Chinese human esophageal cancer.
迁移抑制因子相关蛋白14(MRP14)是一种钙结合蛋白,也被称为S100A9。MRP14与其伴侣MRP8(S100A8)形成的异二聚体分子是中性粒细胞中的主要脂肪酸载体。MRP8/14复合物还参与角质形成细胞中花生四烯酸及其前体的细胞内转运。我们在此展示了MRP14与人类食管癌的关系。在一项初步研究中,运用mRNA差异显示逆转录聚合酶链反应(DD-PCR)对两个食管癌、一个食管腺癌以及匹配的正常相邻黏膜进行检测。用任意引物OPA3进行DD-PCR显示,一条cDNA条带在正常组织中高表达,但在肿瘤对应组织中消失或显著减少。后来鉴定该条带为迁移抑制因子相关蛋白14(MRP14)的3'端。Northern印迹、RT-PCR和Western印迹证实,与食管相邻正常上皮相比,58/64例鳞状细胞癌和2/2例腺癌中MRP14表达下调。在3/3个食管癌细胞系中未检测到MRP14。免疫化学显示MRP14的表达仅限于正常食管上皮。通过PCR和直接DNA测序在MRP14基因的基因组DNA中未发现突变。我们的研究结果表明,MRP14表达降低在中国人类食管癌中是常见现象。