The point mutation in the promoter region and the single nucleotide polymorphism in exon 1 of the cytokeratin 19 gene in human lung cancer cell lines.
The point mutation in the promoter region and the single nucleotide polymorphism in exon 1 of the cytokeratin 19 gene in human lung cancer cell lines.
复制标题
人肺癌细胞系细胞角蛋白19基因启动子区点突变及外显子1单核苷酸多态性。
DOI：
10.1016/s0169-5002(01)00271-9
复制
发表时间：
2001
期刊：
影响因子：
5.3
通讯作者：
T. Ishida
中科院分区：
文献类型：
--
作者：
J. Fujita;K. Dohmoto;S. Hojo;Y. Ueda;S. Bandoh;Y. Yang;Y. Yamaji;Y. Ohtsuki;T. Ishida
检测细胞角蛋白19(CK19)片段的Cyfra 21-1法被广泛用作肺癌的肿瘤标志物。我们曾提出外显子1的PCR扩增失败与CK19基因不能表达密切相关，并假设外显子1内可能存在点突变。为了证明这一点，我们对14个人肺癌细胞系的启动子区域和外显子1进行了序列分析。在14个肺癌细胞系中，有两个细胞系(Lu135和HI1017)的CK19基因启动子区域(−99，G→C)存在点突变。此外，在LU135、HI1017和LC2/AD细胞系中也发现了外显子1(90，T→C，Ala→Ala和179，G→C，Gly→Ala)的点突变。CK19基因启动子区域(位于−99)和外显子1(位于第179位)的点突变通过特定的限制性内切酶分析得到证实。由于在正常志愿者基因组中观察到外显子1(179位)存在相同的点突变，该突变被认为是单核苷酸多态。相比之下，在正常志愿者的基因组中，外显子1的启动子区域没有突变。经过计算机搜索，发现有几个转录因子结合到为扩增外显子1而设计的正义引物序列上。此外，在启动子区域内发生点突变后(−99)，出现了与已知转录因子(AP2)结合的新序列。综上所述，对CK19基因基因组DNA的分析表明，CK19基因的mRNA表达受多种转录因子的调控，这些转录因子与CK19基因启动子区域的特定序列结合。CK19基因启动子区域的突变也下调了CK19的mRNA表达。
The CYFRA 21-1 assay which detects the cytokeratin 19 (CK19) fragment is widely used as a tumor marker for lung cancer. We previously suggested that the failure of PCR amplification of exon 1 is closely related to the inability of the expression of mRNA for CK19, and hypothesized that point mutations might exist within exon 1. In order to prove this, sequence analysis of the promoter region and exon 1 was performed in 14 human lung cancer cell lines. Among the 14 lung cancer cell lines evaluated, point mutations within the promoter region (at −99, G→C) of the CK19 gene were demonstrated in two cell lines (Lu135 and HI1017). In addition, point mutations within exon 1 (at 90, T→C, Ala→Ala and at 179, G→C, Gly→Ala) were also demonstrated in three cell lines (LU135, HI1017, and LC2/AD). Point mutations within the promoter region of CK19 (at −99) and within exon 1 (at 179) were confirmed by analysis of digestion by specific restriction enzymes. Since the same point mutation within exon 1 (at 179) was observed in genomes of normal volunteers, this mutation was considered as a single nucleotide polymorphism. In contrast, there were no mutations within the promoter region of exon 1 in genomes of normal volunteers. After a computer search, it was demonstrated that several transcription factors bind to the sense primer sequence which was designed for amplification of exon 1. In addition, after point mutations within the promoter region occurred (at −99), new sequences appeared to which known transcription factors (AP2) bind. In conclusion, analysis of genomic DNA for CK19 suggested that expression of mRNA for CK19 was regulated by several transcription factors which bound to the specific sequence with the promoter region of the CK19 gene. It was also suggested that the mutation in the promoter region of the CK19 gene down-regulated the expression of mRNA for CK19.
DOI：
10.1074/jbc.m004013200
发表时间：
2000-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
F. Brembeck;Anil K. Rustgi
通讯作者：
F. Brembeck;Anil K. Rustgi
DOI：
10.1006/bbrc.2000.2848
发表时间：
2000-06-16
影响因子：
3.1
作者：
Jiang, JG;DeFrances, MC;Zarnegar, R
通讯作者：
Zarnegar, R
通过含有 AP1 位点的 3 增强子激活角蛋白 19 基因表达。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hu,L;Gudas,LJ
通讯作者：
Gudas,LJ