环境致癌物黄曲霉毒素B1转化大鼠肝干细胞的表达谱分析-生物分子工程专业论文.docx

ABSTRACT Environmental pollutant Aflatoxin B1 (AFB1) is worldspread in nature. Long-term exposure to Aflatoxin B1 (AFB1) often leads to liver cancer. There we focused on the rat hepatic stem-like cell line WB-F344, which was transformed malignantly by treatment with AFB1, on the basis of the cancer stem cell hypothesis. Significantly, the application of gene chip technology to profile the transcriptome in transformed cells could illuminate principle points on mechanism of drug-induced carcinogenesis through the perspective of perturbation the integrated pathway. The rat oval cells WB-F344 were treated by AFB1 and that resulted in inducible malignant transformation. Many transforming focuses appeared. Cells had the ability of anchorage independent growth, and the clone formation ratios were 0.8%, 1.3% and 2%, respectively. Special genes of liver stem cells, Yp, Vimentin, AFP and CK18, were all expressed significantly. We used the Rat OneArray genome microarray to detect the gene expression of WB-F344 cells with the treatment of AFB1. There are 1315 differentially expressed genes (DEGs), and down-regulated genes played a key role. Clustering analysis of DEGs revealed that the effect on cells during the treatment of med- or high-dose AFB1 was conformable. In the whole, up- or down-gene expression in all dosages was superior in DEGs. Gene Ontology (GO) analysis showed that malignant transformation of the liver stem cells was closely correlated to biological process, primary focusing on cell migration, cell adhesion and inflammatory response, etc. Pathway analysis showed that significant pathway mainly focused on MAPK signaling pathway, pathways in cancer, focal adhesion, regulation of actin cytoskeleton, etc. It further indicated that the capital genes in these pathways greatly contributed to the malignant transformation. In DEG interaction network, we found that these genes (MET, CCL2, ITGA1, EGFR, COL1A1, ELN, etc.) were in core. It suggested that these genes might be