Liu Lian-Xin, Jiang Hong-Chi, Liu Zhi-Hua, Zhu An-Long, Zhang Wei-Hui, Wu Lin-Feng, Zhou Jing, Wang Xiu-Qin, Wu Min
Department of Surgery, the First Clinical College, Harbin Medical University, Nangang District, Harbin 150001, Heilongjiang province, PR China.
Oncol Rep. 2003 Nov-Dec;10(6):1771-5.
We investigated the gene expression of the cell cycle/growth regulators in hepatocellular carcinoma (HCC) through the usage of Atlas human cancer array membranes, semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Northern blot. Hybridization of cDNA array membrane was performed with alpha-32P-labeled cDNA probes synthesized from RNA, isolated from HCC and adjacent non-cirrhotic normal liver. RT-PCR of 24 paired specimens and Northern blot of 4 paired specimens were used to confirm the expression patterns of the cell cycle/growth regulator genes identified by Atlas array hybridization. Among 79 genes related to cell cycle/growth regulators, transcription factor DP2 (TFDP-2) and E2F-3 were up-regulated, whereas dual-specificity mitogen-activated protein kinase kinase 1 (MAPKK1) and cell division protein kinase 3 (CDK3) were down-regulated in HCC. RT-PCR of TFDP-2 gave result consistent with Atlas human cancer cDNA array findings. Northern blot analysis of TFDP-2 and E2F-3 of 4 paired specimens all showed up-regulation in HCC compared to normal liver tissues. The results obtained from Atlas microarray provided for the first time a liver cancer-specific expression profile, which identified the gene expressions comprehensively and systematically. The findings may lead to better understanding of the mechanism of onset and progression of HCC. The rapid and high-throughout method of profiling gene expression by cDNA array provides an overview of the key factors that may be involved in HCC. Some genes are reported for the first time in HCC.
我们通过使用阿特拉斯人类癌症阵列膜、半定量逆转录聚合酶链反应(RT-PCR)和Northern印迹法,研究了肝细胞癌(HCC)中细胞周期/生长调节因子的基因表达。用从HCC和相邻的非肝硬化正常肝脏中分离的RNA合成的α-32P标记的cDNA探针进行cDNA阵列膜的杂交。使用24对标本的RT-PCR和4对标本的Northern印迹法来确认通过阿特拉斯阵列杂交鉴定的细胞周期/生长调节因子基因的表达模式。在79个与细胞周期/生长调节因子相关的基因中,转录因子DP2(TFDP-2)和E2F-3在HCC中上调,而双特异性丝裂原活化蛋白激酶激酶1(MAPKK1)和细胞分裂蛋白激酶3(CDK3)在HCC中下调。TFDP-2的RT-PCR结果与阿特拉斯人类癌症cDNA阵列的结果一致。4对标本中TFDP-2和E2F-3的Northern印迹分析均显示,与正常肝组织相比,HCC中它们上调。从阿特拉斯微阵列获得的结果首次提供了肝癌特异性表达谱,该谱全面且系统地鉴定了基因表达。这些发现可能有助于更好地理解HCC的发生和发展机制。通过cDNA阵列进行基因表达谱分析的快速且高通量方法提供了可能参与HCC的关键因素的概述。一些基因在HCC中是首次报道。