Yang Lian-Yue, Wang Wei, Peng Ji-Xiang, Yang Jie-Quan, Huang Gen-Wen
Liver Cancer Laboratory, Department of Surgery, Xiangya Hospital, Changsha 410008, Hunan Province, China.
World J Gastroenterol. 2004 Dec 15;10(24):3569-73. doi: 10.3748/wjg.v10.i24.3569.
To study the difference in gene expression between solitary large hepatocellular carcinoma (SLHCC) and nodular hepatocellular carcinoma (NHCC).
Polymerase chain reaction (PCR) products of 8464 human genes were spotted on a chip in array. DNAs were then fixed on a glass plate. Total RNA was isolated from freshly excised human SLHCC (n = 7) and NHCC (n = 15) tissues, and was reversely transcribed to cDNAs with the incorporation of fluorescent dUTP for preparation of hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After highly stringent washing, cDNA microarray was scanned for the fluorescent signals to display the difference between the two kinds of HCC. In addition, the expression of RhoC and protocadherin LKC was also detected with the reverse transcriptase polymerase chain reaction (RT-PCR) method.
Among the 8464 human genes, 668 (7.89%) genes were expressed differentially at the mRNA levels between SLHCC and NHCC. Three hundred and fifty five (4.19%) genes, including protocadherin LKC, were up-regulated, whereas 313 (3.70%) genes, including RhoC, were down-regulated. The mRNA expression levels of RhoC and protocadherin LKC were confirmed by RT-PCR. Analysis of differentially expressed genes confirmed that our molecular data obtained by cDNA microarray were consistent with the published biochemical and clinical observations of SLHCC and NHCC.
cDNA microarray is an effective technique in screening the difference in gene expression between SLHCC and NHCC. Many of these differentially expressed genes are involved in the invasion and metastasis of HCC. Further analysis of these genes will help to understand the different molecular mechanisms of SLHCC and NHCC.
研究孤立性大肝细胞癌（SLHCC）与结节性肝细胞癌（NHCC）之间的基因表达差异。
将8464个人类基因的聚合酶链反应（PCR）产物点样于芯片上制成阵列。然后将DNA固定在玻璃板上。从新鲜切除的人类SLHCC组织（n = 7）和NHCC组织（n = 15）中提取总RNA，并在掺入荧光dUTP的情况下反转录成cDNA，用于制备杂交探针。然后将混合探针与cDNA微阵列杂交。经过高度严格洗涤后，扫描cDNA微阵列以检测荧光信号，以显示两种肝癌之间的差异。此外，还采用逆转录聚合酶链反应（RT-PCR）方法检测RhoC和原钙黏蛋白LKC的表达。
在8464个人类基因中，有668个（7.89%）基因在SLHCC和NHCC之间的mRNA水平上表达存在差异。包括原钙黏蛋白LKC在内的355个（4.19%）基因上调，而包括RhoC在内的313个（3.70%）基因下调。RT-PCR证实了RhoC和原钙黏蛋白LKC的mRNA表达水平。对差异表达基因的分析证实，我们通过cDNA微阵列获得的分子数据与已发表的关于SLHCC和NHCC的生化及临床观察结果一致。
cDNA微阵列是筛选SLHCC和NHCC之间基因表达差异的有效技术。许多这些差异表达基因参与了肝癌的侵袭和转移。对这些基因的进一步分析将有助于了解SLHCC和NHCC不同的分子机制。
