Nghiên cứu quy định gen mới gây bởi phơi nhiễm liều lượng thấp của Benzo[A]pyrene trong các tế bào Jurkat cells bằng việc sử dụng lai ghép hóa loại trừ ức chế

Aims of this study are to investigate the differential gene expression caused by a carcinogen. the hypothesis is that by employing suppressive subtractive hybridization PCR (SSH PCR), we may identify novel rare up- and down-regulated transcripts that are missed in conventional microarray and proteomic analyses. the authors exposed a non-cytotoxic dose of benzo[a]pyrene (BaP) . to AhR-deficient Jurkat cells and investigated the cellular response in genome-wide gene expression. After treating the cells with 2.5 pM BaP for 48h, the authors isolated mRNA for SSH PCR analysis. the authors screened approximately 15,000 clones to obtain a final of 41 positive clones (20 up- and 21 down-regulated clones). The use of this approach to identify the regulated transcripts which was validated by real-time qPCR. Among these 41 transcripts, the authors observed that 27 of them are related to cell growth, DNA repair, and gene regulation; 3 of them (TCF7, DOCK2, and ZAP70) are related to immune response. Two ot'the down-regulated transcripts BCL2 and TCF7 have been reported to be also down-regulated by cigarette smoke and the rest are novel BaP-regulated transcripts. the data suggested that Jurkat cells are sensitive to DNA damage at a non-cytotoxic dose of BaP because of Jurkat cells down-regulate genes (SART1, SNRK, HIRA, and BCL2) that promote cell growth and up-regulate DNA repair related genes (GTF2H2 and RNASEH1). In addition, immune response is suppressed even though no apparent change in morphology has been observed when comparing the BaP-treated cells with the untreated cells.