The Study On Differential Gene Expression Profiling In L02 Cell Exposed To AFB1

Objective: The present study aimed to explore the miRNA expression profiling to find the AFB1-specific miRNA, and explicit the mechanism of AFB1 in the carcinogenesis the liver cancer; Establish a novel approach for screening genetic targets of genotoxic carcinogen based on GINI (Gene identification by NMD inhibition) and gene chip techniques and to detect differential gene expression profiling in L02 cell exposed to AFB1.Methods: The existence of CYP1A2 enzyme which is essential for the metabolic activation of AFB1 was initially determined by RT-PCR technique. Human liver L02 cells were treated with AFB1 (1μg/ml) for 1 week, followed by soft agar clone formation test to confirm malignant transformation. Gene chip technique was used to detect differential gene expression profiling of micro RNA in L02 cells treated and not treated with AFB1. Based on the principles of GINI technique, the NMD self-repair pathway of transformed cells was blocked by emetine. Gene chip technique was used to detect differential gene expression profiling in transformed L02 cells with and without treatment of emetine.Result: The results of PR-PCR suggested the expression of CYP1A2 in L02 cells. Results of deferential gene expression profiling of micro RNA showed that the mainly up-regulated gene was hsa-miR-671-5p and the mainly down-regulated gene was hsa-miR-96. These genes were close related to protein synthesis, immune-system and psychological functions of cells. The TCS1 is up-expression highly in the cell which is treated with emetine. They have relation with cancer.Conclusion: AFB1 could induce malignant transformation of human liver L02 cells, which may lead to changes in the gene expression. The target genes of miRNA were close related to protein synthesis, immune-system and psychological functions of cells, and thus play an important role in the process of carcinogenesis. GINI technique may be an effective and quick approach to detect target genes of carcinogens. The candidate genes should be confirmed with RT-PCR or North-Blottling in the further research.