Wang Qi-Ming, Jia Lian-Qun, Zhou Hong-Ying, Li Yun-Sheng
Department of Anatomy, Tianjin Medical University, Tianjin, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Oct;23(10):926-8.
To explore the possible mechanism of action of Wnt/beta-catenin signaling pathway in hepatocarcinogenesis by investigating the different expressions of the main members on this signaling pathway in hepatocellular carcinoma cell line HepG2 and L02 cell line.
The mRNAs of Wnt1, Wnt4, beta-catenin, cyclinD1 and c-myc genes were amplified by means of semiquantitative reverse transcription polymerase chain reaction (RT-PCR) in normal liver cell line L02 and hepatocellular carcinoma cell line HepG2, respectively. At the same time, the proteins expression of beta-catenin which was the key member in the Wnt/beta-catenin signaling pathway was examined by immunocytochemical method and Western blot technique.
In normal liver cell line L02, the mRNAs of Wnt1, Wnt4, cyclin D1 and c-myc genes were not detected except for the gene of beta-catenin. In hepatocellular carcinoma cell line HepG2, the mRNAs of Wnt1, beta-catenin, cyclin D1 and c-myc genes were detected except for the gene of Wnt4. Meanwhile, found that beta-catenin proteins were accumulated in the cytoplasm and/or nucleus in HepG2 but only in cell membrane in L02.Using Western blot technique, found that beta-catenin proteins expression was higher in HepG2 than in L02.
The Wnt/beta-catenin signaling transduction pathway is activated with aberrant expression of Wnt1 in hepatocellular carcinoma cell line HepG2.
通过研究Wnt/β-连环蛋白信号通路主要成员在肝癌细胞系HepG2和正常肝细胞系L02中的不同表达情况，探讨该信号通路在肝癌发生中的可能作用机制。
分别采用半定量逆转录聚合酶链反应（RT-PCR）扩增正常肝细胞系L02和肝癌细胞系HepG2中Wnt1、Wnt4、β-连环蛋白、细胞周期蛋白D1和c-myc基因的mRNA。同时，采用免疫细胞化学法和蛋白质印迹技术检测Wnt/β-连环蛋白信号通路关键成员β-连环蛋白的蛋白表达。
在正常肝细胞系L02中，除β-连环蛋白基因外，未检测到Wnt1、Wnt4、细胞周期蛋白D1和c-myc基因的mRNA。在肝癌细胞系HepG2中，除Wnt4基因外，检测到Wnt1、β-连环蛋白、细胞周期蛋白D1和c-myc基因的mRNA。同时发现，β-连环蛋白在HepG2细胞的细胞质和/或细胞核中积累，而在L02细胞中仅存在于细胞膜。采用蛋白质印迹技术发现，HepG2细胞中β-连环蛋白的蛋白表达高于L02细胞。
肝癌细胞系HepG2中Wnt1异常表达激活了Wnt/β-连环蛋白信号转导通路。
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