Positional cloning of the pulmonary adenoma resistance 2 (Par2) locus of the mouse

小鼠肺腺瘤耐药 2 (Par2) 基因座的定位克隆

• 批准号: 14370083
• 负责人: LEE Gang-hong
• 金额: $ 8.51万
• 依托单位: Okinaka Memorial Institute for Medical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370083/
• 关键词: chimeric mouse; lung tumor; resistance locus; chromosome; cell autonomy; candidate gene; urethane; 肺発癌; 耐性遺伝子; 候補遺伝子; Par2; Polι; BALB; c; 129; 3T3; 形質転換; 発がん感受性; 肺腫瘍; ウレタン; ポジショナルクローニング

The Pulmonary adenoma resistance 2 (Par2) locus of the BALB/cByJ mouse, located within 0.5 cM of chromosome 18, is responsible for reducing the mean multiplicity of urethane-induced lung tumors relative to those in C57BL/6J, A/J and C3H/HeJ mice. Thus, BALB/B6-Par2 congenic strain genetically identical to BALB/cByJ except carrying C57BL/6J Par2 alleles develops 7 times more tumors than BALB/cByJ. To gain clues for identification of Par2 candidate genes, we analyzed lung tumorigenesis in BALB/cByJ ⇔ BALB.B6-Par2 chimeric animals. Of 100 tumors induced by urethane in 16 chimeras, 82 originated from BALB.B6-Par2 cells, indicating the Par2 phenotype to be cell-autonomous. In addition, the BALB.B6-Par2- and BALB/cByJ-derived tumors were similar in mean size, implying that the phenotype is primarily expressed during initiation rather than in the promotion stage of carcinogenesis. Given these results, we surveyed a comprehensive mouse genome database and physically mapped Par2 within a 2.3 Mb … More p segment containing three known genes, Poli, Mbd2 and Dcc. Among those, the Poli seemed to be the most reasonable Par2 candidate, since it encodes an extremely error-prone DNA polymerase preferentially incorporating G or T opposite template T in vitro, reminiscent of the Kras2 activation due to an A to G or T point mutation within codon 61 with which most urethane-induced lung tumors are initiated. Indeed, our sequencing of Poli cDNAs from BALB/cByJ, C57BL/6J, A/J and C3H/HeJ lungs revealed 21BALB/cByJ-specific single nucleotide polymorphisms in the coding region accompanied by 7 amino acid substitutions and an elevated frequency of alternative splicing, while no polymorphisms associated with tumor susceptibility were found for either Mbd2 or Dcc. Notably, we obtained evidence that BALB/cByJ Par2 alleles may selectively decrease the frequency of kras2-mutated tumors compared with C57BL/6J alleles. Consequently, the Poli is an intriguing Par2 candidate clearly deserving further evaluation. Less

BALB/cByJ小鼠的肺腺瘤耐药2(PAR2)基因座位于18号染色体的0.5 cM范围内，与C57BL/6J、A/J和C3H/HeJ小鼠相比，PAR2基因座负责降低乌拉坦诱发的肺癌的平均多样性。因此，除携带C57BL/6J PAR2等位基因外，BALB/B6-PAR2同基因株与BALB/cByJ基因相同，发生肿瘤的几率是BALB/cByJ的7倍。为了获得鉴定PAR2候选基因的线索，我们分析了BALB/cByJ⇔BALB.B6-PAR2嵌合动物的肺肿瘤发生。在乌拉坦诱导的16个嵌合体的100个肿瘤中，82个来自BALB.B6-PAR2细胞，表明PAR2的表型是细胞自主的。此外，BALB.B6-PAR2-和BALB/cByJ来源的肿瘤在平均大小上相似，这意味着该表型主要在癌变的起始阶段而不是在癌变的促进阶段表达。鉴于这些结果，我们调查了一个全面的小鼠基因组数据库，并在2.3Mb的…中物理定位了PAR2含有Poli、Mbd2和Dcc三个已知基因的p片段较多。其中，PolI似乎是最合理的PAR2候选序列，因为它编码一种极易出错的DNA聚合酶，在体外优先结合G或T相反的模板T，这让人想起Kras2的激活，这是由于61号密码子的A到G或T点突变导致的，大多数乌拉坦诱导的肺肿瘤都是由61号密码子引发的。事实上，我们对BALB/cByJ、C57BL/6J、A/J和C3H/HeJ肺的Poli cDNA进行了测序，发现编码区有21BALB/cByJ特异的单核苷酸多态，伴随着7个氨基酸替换和选择性剪接频率增加，而Mbd2和DCC没有发现与肿瘤易感性相关的多态。值得注意的是，我们获得的证据表明，与C57BL/6J等位基因相比，BALB/cByJ PAR2等位基因可能选择性地降低Kras2突变肿瘤的频率。因此，POLI是一个耐人寻味的PAR2候选方案，显然值得进一步评估。较少

项目成果

Lee, G.-H., et al.: "Analysis of lung tumorigenesis in chimeric mice indicates the Pulmonary adenoma resistance 2 (Par2) locus to operate in the tumor-initiation stage in a cell-aulonomous manner : detection of polymorphisms in the Polι gene as a candidat

Lee, G.-H. 等人：“对嵌合小鼠肺肿瘤发生的分析表明，肺腺瘤耐药 2 (Par2) 位点以细胞自主方式在肿瘤起始阶段发挥作用：检测肺腺瘤耐药性 2 (Par2) 基因座中的多态性Polι基因作为候选者

Genetic linkage between Poli deficiency and increased susceptibility to lung tumors in mice.

Poli 缺乏与小鼠肺部肿瘤易感性增加之间的遗传联系。

• 期刊: Cancer Science (発表予定)
• 作者: Lee;G-H;Matsushita;H.
• 通讯作者: H.

Genetic linkage between Polι deficiency and increased susceptibility to lung tumors in mice.

Polι缺乏与小鼠肺部肿瘤易感性增加之间的遗传联系。

• 期刊: Cancer Science (印刷中)
• 作者: Lee;G-H;Matsushita;H
• 通讯作者: H

Analysis of lung tumorigenesis in chimeric mice indicates the Pulmonary adenoma resistance 2(Par2) locus to operate in the tumor-initiation stage in a cell-autonomous manner : detection of polymorphisms in the Poli gene as a candidate for Par2.

对嵌合小鼠肺肿瘤发生的分析表明，肺腺瘤抗性 2 (Par2) 位点以细胞自主方式在肿瘤起始阶段发挥作用：检测作为 Par2 候选者的 Poli 基因中的多态性。

• 发表时间: 2003
• 期刊: Oncogene 22
• 作者: Lee;G.-H.;Nishimori;H.;Sasaki;Y.;Matsushita;H.;Kitagawa;T.;Tokino;T.
• 通讯作者: T.

Gang-Hong Lee et al.: "Analysis of lung tumorigenesis in chimeric mice indicates the Pulmonary adenoma resistance 2 (Par2) locus to operate in the tumor-initiation stage in a cell-autonomous manner : detection of polymorphisms in the Polι gene as a candid

Gang-Hong Lee 等人：“对嵌合小鼠肺肿瘤发生的分析表明，肺腺瘤耐药 2 (Par2) 位点以细胞自主方式在肿瘤起始阶段发挥作用：检测 Polι 基因中的多态性作为坦率