Kidney-targeted Birt-Hogg-Dube gene inactivation in a mouse model: Erk1/2 and Akt-mTOR activation, cell hyperproliferation, and polycystic kidneys
Kidney-targeted Birt-Hogg-Dube gene inactivation in a mouse model: Erk1/2 and Akt-mTOR activation, cell hyperproliferation, and polycystic kidneys
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DOI：
10.1093/jnci/djm288
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发表时间：
2008-01-16
影响因子：
10.3
通讯作者：
Schmidt, Laura S.
中科院分区：
文献类型：
--
作者：
Baba, Masaya;Furihata, Mutsuo;Schmidt, Laura S.
Birt-Hogg-Dube（BHD）综合征患者的BHD肿瘤抑制基因存在生殖系突变，与肾癌风险增加相关。BHD编码卵泡素，一种蛋白质，可能与能量和营养敏感的5 '-AMP激活的蛋白激酶-哺乳动物雷帕霉素靶（AMPK-mTOR）signalingpathways.Methods相互作用，我们使用重组工程方法产生小鼠与条件BHD等位基因，并引入钙粘蛋白16（KSP）-Cre转基因靶向BHD失活的肾脏。通过BrdU掺入和磷酸化组蛋白H3染色来测量肾细胞增殖。采用Wilcoxon秩和、Student t和Welch t检验分析肾脏重量数据。对小鼠肾细胞和组织进行苏木精和伊红染色和免疫印迹分析以及细胞周期和信号蛋白的免疫组化。用mTOR抑制剂雷帕霉素处理BHD敲除小鼠和从BHD敲除小鼠和对照小鼠分离的肾细胞。通过Kaplan-Meier分析评价小鼠存活率。结果BHD基因敲除小鼠出现了增大的多囊肾，并在3周龄时死于肾衰竭。靶向BHD敲除导致肾脏中Raf-extracellular signal-regulated protein kinase（Erk）1/ 2和Akt-mTOR途径的激活，并增加细胞周期蛋白的表达和细胞增殖。雷帕霉素处理的BHD敲除小鼠的肾脏比缓冲液处理的BHD敲除小鼠的肾脏小（每组n = 4 - 6只小鼠，相对肾脏/体重比，平均值= 4.64%对12.2%，差异= 7.6%，95%置信区间= 5.2%至10.0%; P
Background Patients with Birt-Hogg-Dube ( BHD) syndrome harbor germline mutations in the BHD tumor suppressor gene that are associated with an increased risk for kidney cancer. BHD encodes folliculin, a protein that may interact with the energy- and nutrient-sensing 5'-AMP-activated protein kinase-mammalian target of rapamycin (AMPK-mTOR) signaling pathways.Methods We used recombineering methods to generate mice with a conditional BHD allele and introduced the cadherin 16 ( KSP)-Cre transgene to target BHD inactivation to the kidney. Kidney cell proliferation was measured by BrdU incorporation and phospho-histone H3 staining. Kidney weight data were analyzed with Wilcoxon's rank-sum, Student's t, and Welch's t tests. Hematoxylin and eosin staining and immunoblot analysis and immunohistochemistry of cell cycle and signaling proteins were performed on mouse kidney cells and tissues. BHD knockout mice and kidney cells isolated from BHD knockout and control mice were treated with the mTOR inhibitor rapamycin. Mouse survival was evaluated by Kaplan-Meier analyses. All statistical tests were two-sided.Results BHD knockout mice developed enlarged polycystic kidneys and died from renal failure by 3 weeks of age. Targeted BHD knockout led to the activation of Raf-extracellular signal-regulated protein kinase ( Erk)1/ 2 and Akt-mTOR pathways in the kidneys and increased expression of cell cycle proteins and cell proliferation. Rapamycin-treated BHD knockout mice had smaller kidneys than buffer-treated BHD knockout mice had ( n = 4 - 6 mice per group, relative kidney/ body weight ratios, mean = 4.64% vs 12.2%, difference = 7.6%, 95% confidence interval = 5.2% to 10.0%; P