Tan Zhijun, Hu Xiangui, Ying Kang, Li Yao, Tang Rong, Cao Guisong, Tang Yan, Jin Gang
Department of General Surgery, Changhai Hospital, Shanghai 200433, China.
Zhonghua Zhong Liu Za Zhi. 2002 May;24(3):243-6.
Analysis of differential gene expression profiles by cDNA microarray in pancreatic carcinoma with or without lymphatic metastasis.
cDNA microarray was prepared by spotting polymerase chain reaction (PCR) products of 4 000 human genes onto specially treated glass slides. The cDNA probes were prepared by labeling normal tissue mRNA and cancer tissue mRNA with Cy3-dUTP and Cy5-dUTP, separately through reverse transcription. The mixed probes were, then, hybridized to the cDNA microarray. The chips were scanned by ScanArray 3000 laser scanner (General Scanning, Inc) on two wavelengths. The acquired image was analyzed by ImaGene 3.0 software (BioDiscovery, Inc). The intensity of each spot on the two wavelengths represented the quantity of Cy3-dUTP and Cy5-dUTP, with Cy5 to Cy3 ratio computed on each.
Fifty-six genes (including 24 previously reported) exhibited differential expressions in 2 specimens of pancreatic carcinoma with lymphatic metastasis and 2 without.
cDNA microarray provides an promising approach to specific gene expressions of the presence of lymphatic metastasis in human pancreatic carcinoma.
通过cDNA微阵列分析有或无淋巴转移的胰腺癌组织中的差异基因表达谱。
通过将4000个人类基因的聚合酶链反应（PCR）产物点样到经过特殊处理的载玻片上来制备cDNA微阵列。分别通过逆转录用Cy3-dUTP和Cy5-dUTP标记正常组织mRNA和癌组织mRNA来制备cDNA探针。然后，将混合探针与cDNA微阵列杂交。用ScanArray 3000激光扫描仪（通用扫描公司）在两个波长下扫描芯片。用ImaGene 3.0软件（生物发现公司）分析采集到的图像。两个波长下每个点的强度代表Cy3-dUTP和Cy5-dUTP的量，并计算每个点的Cy5与Cy3的比值。
在2例有淋巴转移和2例无淋巴转移的胰腺癌标本中，有56个基因（包括24个先前报道的基因）表现出差异表达。
cDNA微阵列为分析人胰腺癌中淋巴转移存在时的特定基因表达提供了一种有前景的方法。
