Cao Xueyuan, Tsukamoto Tetsuya, Nozaki Koji, Mizoshita Tsutomu, Ogasawara Naotaka, Tanaka Harunari, Takenaka Yoshiharu, Kaminishi Michio, Tatematsu Masae
Division of Oncological Pathology, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya, Aichi 464-8681, Japan.
Cancer Sci. 2004 Jun;95(6):487-90. doi: 10.1111/j.1349-7006.2004.tb03237.x.
The goal of this study was to elucidate whether beta-catenin gene mutations might contribute to glandular stomach carcinogenesis in Helicobacter pylori (H.pylori)-infected Mongolian gerbils. Firstly, exon 3 of gerbil beta-catenin cDNA, a mutation hot spot, was cloned and sequenced and found to have 89.3% homology with the human form and 95.5% with the rat and mouse forms. Peptide sequence in this region was shown to be 100% conserved in these mammals. Then, 45 stomach adenocarcinomas induced with N-methyl-N-nitrosourea (MNU) plus H. pylori infection and 7 induced with MNU alone were examined for beta-catenin expression by immunohistochemistry and for DNA mutations using a combination of microdissection and PCR-single strand conformation polymorphism analysis. One gastric cancer in the MNU + H. pylori group (2.2%) displayed nuclear (N) beta-catenin localization, 3 (6.7%) showed cytoplasmic (C) distribution in local regions, and 41 (91.1%) demonstrated cell membrane (M) localization. Tumors induced by MNU alone showed only membranous beta-catenin localization (7/7). Analysis of exon 3 of the beta-catenin gene dem-onstrated all tumors with membrane or cytoplasmic staining as well as surrounding normal mucosa (S) to feature wild-type beta-catenin. In contrast, the lesion with nuclear staining had a missense mutation at codon 34 [GAC (Gly) --> GAA (Glu)] in exon 3 (1/1 = 100%, N vs. M, P < 0.05; and N vs. S, P < 0.05). In conclusion, these results suggest that beta-catenin may not be a frequent target for mutation in stomach carcinogenesis in MNU + H. pylori-treated gerbils.
本研究的目的是阐明β-连环蛋白基因突变是否可能在幽门螺杆菌(H.pylori)感染的蒙古沙鼠腺胃癌发生过程中发挥作用。首先,克隆并测序了沙鼠β-连环蛋白cDNA的第3外显子(一个突变热点),发现其与人类序列的同源性为89.3%,与大鼠和小鼠序列的同源性为95.5%。该区域的肽序列在这些哺乳动物中显示为100%保守。然后,通过免疫组织化学检测了45例经N-甲基-N-亚硝基脲(MNU)加H.pylori感染诱导的胃腺癌以及7例仅经MNU诱导的胃腺癌中β-连环蛋白的表达情况,并使用显微切割和PCR-单链构象多态性分析相结合的方法检测了DNA突变。MNU + H.pylori组中的1例胃癌(2.2%)显示β-连环蛋白定位于细胞核(N),3例(6.7%)在局部区域显示为细胞质(C)分布,41例(91.1%)表现为细胞膜(M)定位。仅经MNU诱导的肿瘤仅显示β-连环蛋白定位于细胞膜(7/7)。对β-连环蛋白基因第3外显子的分析表明,所有显示膜或细胞质染色的肿瘤以及周围正常黏膜(S)均具有野生型β-连环蛋白特征。相反,细胞核染色的病变在外显子3的第34密码子处发生了错义突变[GAC(甘氨酸)→GAA(谷氨酸)](1/1 = 100%,N与M相比,P < 0.05;N与S相比,P < 0.05)。总之,这些结果表明,在MNU + H.pylori处理的沙鼠胃癌发生过程中,β-连环蛋白可能不是常见的突变靶点。