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食管癌细胞中的错配修复与微卫星不稳定性

Mismatch repair and microsatellite instability in esophageal cancer cells.

作者信息

Uchida N, Kumimoto H, Nishizawa K, Tokumasu S, Harada H, Shimada Y, Ishizaki K

机构信息

Central Laboratory and Radiation Biology, Aichi Cancer Center Research Institute, Nagoya, Japan.

出版信息

Int J Cancer. 2001 Mar 1;91(5):687-91.

Abstract

Using in vitro mismatch repair (MMR) assay, we have identified 3 of 22 esophageal cancer cell lines exhibiting reduced MMR activity. By means of gel-shift assay, decreased binding ability to GT mismatch and CA loop was observed in these 3 cell lines. However, we could not find any mutations in the hMSH2, hMSH3 and hMSH6 genes, the protein products of which exhibit mismatch binding activity in human cells. In addition, when using antibodies against 5 MMR-related proteins (hMSH2, hMSH3, hMSH6, hPMS2 and hMLH1), no aberrant expression was detected in any of them. When we examined 9 microsatellite loci in endogenous genomic DNA, these 3 esophageal cancer cell lines, deficient in MMR, did not exhibit microsatellite instability. However, when we examined the repetitious sequence on exogenous plasmid DNA which was introduced into these 3 esophageal cancer cells, the results suggested that MMR deficiency in esophageal cancer cells could result in moderate instability of the exogenous sequence.

摘要

通过体外错配修复(MMR)检测,我们在22种食管癌细胞系中鉴定出3种错配修复活性降低的细胞系。通过凝胶迁移试验,在这3种细胞系中观察到与GT错配和CA环的结合能力下降。然而,我们在hMSH2、hMSH3和hMSH6基因中未发现任何突变,这些基因的蛋白质产物在人类细胞中表现出错配结合活性。此外,当使用针对5种MMR相关蛋白(hMSH2、hMSH3、hMSH6、hPMS2和hMLH1)的抗体时,未在其中任何一种中检测到异常表达。当我们检测内源性基因组DNA中的9个微卫星位点时,这3种错配修复缺陷的食管癌细胞系未表现出微卫星不稳定性。然而,当我们检测导入这3种食管癌细胞的外源性质粒DNA上的重复序列时,结果表明食管癌细胞中的错配修复缺陷可能导致外源性序列的中度不稳定性。

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