Guo Kun, Li Yan, Kang Xiaonan, Sun Lu, Cui Jiefeng, Gao Dongmei, Liu Yinkun
Liver Cancer Institute, Zhongshan Hospital, Fudan University, No. 180 Fenglin Road, Shanghai, China.
Clin Exp Metastasis. 2009;26(3):189-95. doi: 10.1007/s10585-008-9230-y. Epub 2008 Dec 27.
Considerable interests have recently been focused on mechanism of human hepatocellular carcinoma (HCC) metastasis-the most fundamental characteristics of HCC and the ultimate cause of most HCC mortality, so screening more potential early prognostic marker and therapeutic target is urgent. In this study, we screened genome of three HCC cell lines with consistently increased metastatic potentials and sharing same genetic background, through DNA microarray and found consecutively up-regulated expression of PKCbeta in these cell lines compared to others PKCs, which was reconfirmed by real time RT-PCR and western blot analysis. Moreover, it was found, after efficient silence of PKCbeta by RNAi assay or inhibition of PKCbeta activity by a specific inhibitor LY317615, migration and invasion of HCC cells significantly decreased. In addition, depletion of PKCbeta protein significantly reversed the enhancement of PMA-stimulated HCC migration and invasion ability in vitro. All the data suggest a key role of PKCbeta in HCC motility and PKCbeta may be a potential therapeutic target.
近年来，相当多的研究兴趣集中在人类肝细胞癌（HCC）转移机制上，这是HCC最基本的特征，也是大多数HCC死亡的最终原因，因此筛选更多潜在的早期预后标志物和治疗靶点迫在眉睫。在本研究中，我们通过DNA微阵列筛选了三种具有持续增加的转移潜能且具有相同遗传背景的HCC细胞系的基因组，发现与其他蛋白激酶C（PKC）相比，这些细胞系中PKCβ的表达持续上调，这通过实时逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹分析得到再次证实。此外，通过RNA干扰试验有效沉默PKCβ或用特异性抑制剂LY317615抑制PKCβ活性后，发现HCC细胞的迁移和侵袭明显减少。另外，PKCβ蛋白的缺失显著逆转了体外佛波酯（PMA）刺激的HCC迁移和侵袭能力的增强。所有数据表明PKCβ在HCC运动中起关键作用，PKCβ可能是一个潜在的治疗靶点。
