基因表达谱芯片技术筛选舌癌转移相关基因

Screening and Identification of Metastasis-related Gene Expression in Tongue Carcinoma with cDNA Mi- croarray Assay

目的应用基因表达谱芯片技术对口腔舌癌细胞系Tca8113及其脑转移细胞株Tb的基因表达谱进行对比分析并筛选差异表达基因,以期从基因水平上探讨舌癌转移机制。方法以舌癌细胞系Tca8113及其脑转移细胞株Tb作为研究肿瘤转移分子机制的模型,提取mRNA,逆转录为cDNA,用Cy3和Cy5荧光染料标记,制备成cDNA 探针,与基因表达谱芯片进行杂交扫描和分析;并应用RT-PCR技术对其中6个基因进行验证。结果在1152个基因表达谱的筛选中,37个基因表达有差异(2倍以上),其中15个基因表达水平上调,22个基因表达水平下调。 RT-PCR技术对其中6个基因表达差异的验证结果与基因芯片结果一致。结论舌癌细胞转移侵袭可能与多个基因相关。

Objective To identify metastasis-associated genes in tongue carcinoma and to better understand the mechanism underlying tongue carcinoma metastasis. To compared mRNA expression profiles of two tongue carcinoma cell strains with high and low metastatic potentials using microarray technology. Methods Tca8113 and Tb cells were used as model systems to study the molecular mechanism of tumor metastasis. Two fluorescent eDNA probes labeled with Cy3 and Cy5 dyes were prepared from the mRNA samples of Tea8113 and Tb cells by reverse transcription method. The two color probes were then mixed and hybridized to the eDNA chip constructed by double dots of 1 152 human genes, and scanned at two wave lengths. Differential expression genes from the above two cell lines were analyzed using computer. Then six of the different expression genes were further validated by RT-PCR technique. Results In the 1152 clones of known genes and expressed sequence tags that were analyzed, 37 showed significantly different （minimum 2 folds） expression levels in two cell lines. Among the 37 genes, 15 were up regulated （with ratio more than 2） and 22 down regulated （with ratio less than 1/2）. The results of RT-PCR analysis were coincident with those of micrearray assay. Conclusion Some of these genes are known to be involved in human tumor antigen, immune surveillance, adhesion, cell signaling pathway and growth control. It is suggested that the micrearray in combination with a relevant analysis facilitates rapid and simultaneous identification of multiple genes of interests and in this study it provided a profound clue to screen candidate targets for early diagnosis and intervention.