癌变·畸变·突变

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甲基苄基亚硝胺诱导大鼠食管癌癌前病变食管组织中β-catenin表达及其Ser675位点磷酸化水平的变化

施文荣,刘  艳*   

  1. 福建中医药大学中西医结合学院,福建  福州  350108
  • 收稿日期:2014-04-04 修回日期:2014-05-28 出版日期:2014-09-30 发布日期:2014-09-30
  • 通讯作者: 刘 艳,E-mail:liu_yan1976@126.com
  • 作者简介:施文荣(1975- ),男,福建省莆田市人,硕士,副教授,研究方向:中西医结合基础研究。
  • 基金资助:

    福建省自然科学基金(2012J01389)

Effects of methyl benzyl nitrosamine on β-catenin and p-β-catenin(Ser675) in esophageal precancerous lesions in Wistar rats

SHI Wen-rong,LIU Yan*   

  1. College of Integrated Traditional Chinese and Western Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350108, Fujian, China
  • Received:2014-04-04 Revised:2014-05-28 Online:2014-09-30 Published:2014-09-30

摘要:

目的: 探讨甲基苄基亚硝胺(MBNA)诱导食管癌癌前病变食管组织中β-连环蛋白(β-catenin)mRNA转录、蛋白表达及其Ser675位点磷酸化水平的变化。方法:Wistar大鼠按3.5 mg/kg皮下注射0.15% MBNA溶液,每周2次作为模型组,以同批次正常大鼠(皮下注射生理盐水)为对照组,每组8只,连续10周,于造模第10周处死2组大鼠,观察食管黏膜病理变化,采用定量PCR检测食管组织中β-catenin mRNA转录水平,Western blot检测β-catenin蛋白及其Ser675位点磷酸化蛋白表达水平,免疫组化检测β-catenin蛋白表达的定位情况。结果:MBNA诱导10周后模型组大鼠均可见食管黏膜粗糙,部分可见乳突瘤,食管黏膜病理学检查呈轻度不典型增生,且模型组大鼠食管组织中β-catenin mRNA转录水平(0.619±0.086)较正常组(1.000±0.235)降低(P<0.01);β-catenin蛋白表达水平(1.476±0.363)较正常组(1.000±0.496)升高(P<0.05);Ser675位点磷酸化β-catenin蛋白表达水平(2.150± 0.469)较正常组(1.000±0.367)升高(P<0.01);免疫组化结果显示β-catenin定位表达于食管黏膜上皮细胞,且模型组中的表达较正常组增强。结论:β-catenin蛋白的异常表达及其Ser675位点磷酸化水平的升高与食管癌变的病理变化可能直接相关,可作为食管癌研究的靶点。

关键词: 甲基苄基亚硝胺, 食管癌, 癌前病变, &beta, -catenin, Ser675位点, 磷酸化

Abstract:

OBJECTIVE: To find the changes of β-catenin and phosphorylated β-catenin (p-β-catenin) Ser675 in esophageal precancerous lesions in Wistar rats induced by methyl benzyl nitrosamine (MBNA). METHODS: Wistar rats received MBNA injections at dose of 3.5 mg/kg 2 times per week. At 10 weeks,the pathological changes of esophageal mucosa were examined. β-catenin mRNA level was measured by quantitative real-time PCR and the expressions of β-catenin protein and p-β-catenin(Ser675) were evaluated by Western blot. The location of β-catenin was detected by immunohistochemistry. RESULTS:Compared with normal group,the level of β-catenin mRNA was significantly decreased,the levels of β-catenin and p-β-catenin (Ser675) protein were significantly increased in treated group. CONCLUSION:The elevated protein levels of β-catenin and p-β-catenin (Ser675) were related with esophageal precancerous lesions in Wistar rats induced by MBNA ,and could be considered as an indicator of esophageal carcinoma.

Key words: methyl benzyl nitrosamine, esophageal carcinoma, precancerous lesions, β-catenin, Ser675, phosphorylation