摘要
AIM: To study the difference in gene expression between solitary large hepatocellular carcinoma (SLHCC) and nodular hepatocellular carcinoma (NHCC).METHODS: 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.RESULTS: Among the 8 464 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 upregulated, whereas 313 (3.70%) genes, including RhoC,were down-regulated. The mRNA expression levels of RhoC and protocadherin LKCwere 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.CONCLUSION: 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.
AIM:To study the difference in gene expression between solitary large hepatocellular carcinoma(SLHCC)and nodular hepatocellular carcinoma(NHCC). METHODS: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 RhoCand protocadherin LKC was also detected with the reverse transcriptase polymerase chain reaction(RT-PCR)method. RESULTS:Among the 8 464 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 LKCwere 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. CONCLUSION: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.
基金
Supported by National Key Technologies RD Program of China during the 10~(th) Five-year plan period,No.2001BA703B04
National Natural Science Foundation of China,No.30371595
Hunan Provin e Developing Planning Committee,No.2001-907
