CDC25B基因在肝细胞肝癌的表达  

Expression of CDC25B gene in hepatocellular carcinoma

作　　者：石欣[1] 高乃荣[1] 卫文俊[1] 程张军[1] 汤永辉[1] 

机构地区：[1]东南大学附属中大医院普外科,南京市210009

出　　处：《中华肝胆外科杂志》2007年第1期38-40,共3页Chinese Journal of Hepatobiliary Surgery

摘　　要：目的探讨CDC25B基因在肝细胞肝癌（HCC）组织中的表达，以探讨其在HCC发生和发展中的作用。方法联合应用激光显微切割和实时定量PCR技术检测24例正常肝脏组织和24例HCC组织中CDC25B基因的表达，PCR产物的定量方法采用比较Ct法；应用Western blot检测其蛋白的表达水平；应用免疫组织化学方法检测增殖细胞核抗原（proliferating cell nuclear antigen，PC—NA）的表达。结果在显微镜下用激光显微切割机，将单个肝细胞成功切下，提取RNA后，逆转录成cDNA，实时定量PCR发现CDC25B基因平均模板量在正常肝脏组织中为0．0006350±0．00021，HCC组织中为0．001988±0．00065，上调约3倍（P〈0．01）。CDC25B mRNA的表达与PCNA明显相关（r=0．45，P〈0．05）。HCC组织中CDC25B蛋白也过度表达，正常肝脏组织CDC25B蛋白表达为1．02±0．43，HCC组织为6．01±1．66，上调约6倍（P〈0．01）。结论CDC25B在HCC组织中表达明显上调，该基因的高表达与细胞周期调节机制的关系以及在HCC细胞的恶性转化中的作用还有待进一步探讨。Objective To investigate the expression of cell division cycle 25 B （CDC25B） gene in hepatocellular carcinoma （HCC） and explore its role in pathogenesis of HCC. Methods The expression of CDC25B gene in 24 cases of HCC and 24 cases of normal liver was determined by laser microdissection and quantitative real-time PCR （QRT-PCR）. The product of QRT-PCR was quantified by comparative Ct method. The protein level of CDC25B was determined by Western blot. Proliferating cell nuclear antigen （PCNA） was evaluated with immunohistochemistry. Results Single cell was microdissected from histological tissue using an ultraviolet laser micromanipulator. Once captured, the RNA could be extracted from the isolated cell（s）, reversely transcribed and used for amplification by QRT-PCIL The average quantity of CDC25B gene in normal liver and HCC detected by QRT-PCR was 0. 000635±0. 00021 and 0. 001988±0. 00065, respectively, with about 3-fold increasing. The expression of CDC25B in HCC was correlated to the expression of PCNA （P〈0. 05）. Western blot revealed that CDC25B protein was 1.02±0. 43 in normal liver and 6.01±1.66 in HCC. Statistical analysis showed that it was also significantly overexpressed in HCC tissue by 6 folds （P〈0. 01）. Conclusions CDC25B is upregulated in HCC. The real relationship between its overexpression and deregulation of cell cycle or malignant transformation of liver cancer cell still remains to be clarified.

关 键 词：癌 肝细胞 CDC25B基因 激光显微切割 实时定量PCR 

分 类 号：R686[医药卫生—骨科学]