[Screening for lymphatic metastasis-associated genes in mouse hepatocarcinoma cell lines Hca-F and Hca-P using gene chip].

ABSTRACT

BACKGROUND & OBJECTIVE: Metastasis is a complex process involving multiple genetic changes. The high mortality and poor prognosis caused by metastasis in malignant tumor patients and the uncertain mechanisms are always the prominent problems in the field of oncology. In order to screen for lymphatic metastasis-associated genes, the gene expression profiles of mouse hepatocarcinoma cell lines Hca-F (highly metastatic) and Hca-P (low metastatic) were compared by gene chip. METHODS: Total RNA was isolated from Hca-F and Hca-P cells, and synthesized into double-stranded cDNA, then synthesized into biotin-labeled cRNA probes by in vitro transcription. The cRNA probes were separately hybridized with Affymetrix GeneChip MOE430A (containing 22,690 transcripts, including 14,500 known mouse genes and 4,371 ESTs), and the signals were scanned by the GeneArray Scanner. The results were analyzed by bioinformatics. RESULTS: Compared with the gene expression profile of Hca-P cells, 901 (6.2%) genes and 129 (3%) ESTs were up-regulated by at least 2 folds in Hca-F cells; 33 genes, including endoglin (EDG; CD105), Mcam (Muc18; Mel-CAM; CD146), Cdc42ep5 (CEP5; Borg3), Ptprr (protein tyrosine phosphatase, receptor type, R), F2r [coagulation factor II (thrombin) receptor; Par1; ThrR], D7Ertd458e (necl-5), NR1D1, Serpin h1 (HSP47), AXL, Mak, and Areg (AR), were up-regulated 13.93-29.86 folds. According to Gene Ontology and Treeview analysis, these 33 genes were involved in angiogenesis, cell adhesion, signal transduction, cell motility, transcription, chaperone activity, protein kinase activity, receptor binding, and so on. CONCLUSION: Many lymphatic metastasis-associated genes were screened by high-throughput gene chip method; validating their cellular functions will help to identify the key or candidate gene/pathway responsible for lymphatic metastasis, which might be used as diagnostic markers and therapeutic targets for lymphatic metastasis.