应用表达芯片筛选小鼠肝癌淋巴道转移相关基因

Identification of lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines using cDNA microarray

目的采用表达芯片比较高、低淋巴道转移力小鼠肝癌腹水型细胞株Hca-F和Hca-P的基因表达谱,以筛选出与肿瘤淋巴道转移相关的基因。方法分别提取Hca-F和Hca-P细胞的总RNA,反转录合成生物素标记的cRNA探针,并与A ffy-m etrix GeneCh ip MOE430A(包括22 690个转录本,对应于约14 500个小鼠已知基因和4 371个EST)杂交,检测结果利用生物信息学进行分析。结果Hca-F和Hca-P相比,在14 500个已知基因中,有901个(6.2%)表达上调幅度≥2倍;在4 371个EST中,有129个(3%)上调幅度≥2倍。公布了差异表达≥8倍的37个基因并根据Gene Ontology(GO)分类和TreeV iew分析,按照其参与的生物学过程和具有的分子功能进行了功能分类。其中有19个基因参与了组织发生、细胞黏附、信号转导、细胞生长、分化及代谢等的生物学过程,29个基因分别具有转运、移动、蛋白激酶、蛋白结合及受体活性等分子功能,7个基因的生物学功能尚不清楚。结论表达芯片检测与肿瘤淋巴道转移模型相结合,为肿瘤转移研究提供新方法、新思路,一些过量表达的基因将为肿瘤转移机制的研究提供新线索。

Purpose To screen lymphatic metastasis-associated genes, and to compare the transcriptional profiles of mouse hapatocarcinoma ascites cell lines Hca-F （ highly metastatic ） and Hca-P （ low metastatic ） using cDNA microarray. Methods Total RNA was isolated from Hca-F and Hca-P cells and synthesized into double-stranded cDNA. In vitro transcription double-stranded cRNA was labeled with biotin （biotin-labeled cRNA as a probe）. The cRNA probes were hybridized with Affymetrix GeneChip MOE430A （containing 22 690 transcripts, including 14 500 known mouse genes and 4 371 ESTs）. The results were analyzed by bioinformatics. Resuits Out of the 14 500 genes and 4 371 ESTs investigated, 901 （6. 2% ） genes and 129（3% ） ESTs were up-regulated to at least 2 fold. The top 37 genes were up-regulated at least 8 fold were presented. According to the Gene Ontology （GO） classification and Tree- View analysis, these 37 genes were classified into differential biological process profiles and molecular function profiles. Among them, 19 genes were involved in the biological process, such as development, cell adhesion, signal transduction, cell growth, cell differentiation and metabolism. 29 genes were correlated with the transporter activity, motor activity, kinase activity, protein binding and receptor activity. The functions of 7 genes were unclear. Conclusions cDNA microarray combined with lymphatic metastasis models might contribute new methods and clues to the tumor lymphatic metastasis research. Some overexpressed genes might provide novel clues to the molecular mechanisms of tumor lymphatic metastasis.