摘要
目的研究苯并[a]芘恶性转化的肺癌细胞16HBE-T中circRNA 001988的表达及其下游靶基因的表达改变,并探索该circRNA在化学致癌过程中可能发挥的作用。方法采用q RT-PCR检测circRNA 001988在16HBE-T中的表达,进一步在人肺癌细胞株A549、H460和H446中检测其表达,同时在20例肺癌组织样品中检测了circRNA 001988的表达水平;利用生物信息学分析软件mi RSystem和RegRNA,预测得到circRNA 001988靶向的microRNA以及与其相互作用的mRNA;采用q RT-PCR在16HBE-T和H460中检测microRNA及mRNA的表达。结果与16HBE相比,circRNA 001988在16HBE-T中的表达下调了(6.54±0.75)倍,差异有统计学意义(P<0.01),而在A549和H460细胞中表达分别下调(2.06±0.38)倍、(3.02±0.51)倍,差异均有统计学意义(P<0.05)。与癌旁组织相比,circRNA 001988在肺癌组织中的表达平均下调了(2.11±0.71)倍,差异有统计学意义(P<0.05)。结合生物信息学预测分析和qRT-PCR检测发现,相对于16HBE,circRNA 001988靶向的hsa-mi R-382-5p和hsa-miR-583在16HBE-T和H460细胞中的表达均上调,而hsa-miR-382-5p和hsa-miR-583共同作用的mRNA(CCDC6)的表达水平分别降低(26.20±3.92)倍、(12.61±1.48)倍。结论 circRNA 001988在苯并[a]芘恶性转化的16HBE-T细胞中的表达水平明显降低,它在苯并[a]芘致肺癌过程中可能发挥抑癌因子的作用,并可能通过内源性竞争的机制影响其下游的microRNA和mRNA的表达。
Objective To investigate the expression of circRNA 001988 and its target genes in 16HBE-T cells that were transformed by benzo[a]pyrene, and to explore the role of this circularRNA in the process of chemical carcinogenesis. Methods The expression of circRNA 001988 in 16HBE-T, A549,H460,H446 cells lines and 20 paired neoplastic and adjacent tissues was measured by quantitative Real-Time PCR（q RT-PCR）; MicroRNAs（miRNAs） targeted by circRNA 001988 and mRNAs interacted with this miRNAs were predicted using bioinformatics analysis software of mi RSystem and RegRNA. MiRNAs and mRNAs were measured by q RT-PCR in 16HBE-T and H460 cells line. Results circRNA 001988 was downregulated by（6.54±0.75） folds in 16HBE-T cell line,（2.06±0.38） folds in A549 cell line and（3.02±0.51） folds in H460 cell line, and the differences were statistically significant（P〈 0.05）. Expression of circRNA 001988 in malignant lung tissues was averagely downregulated by（2.11±0.71）folds compared with para-carcinoma tissues, and the differences were statistically significant（P0.05）. Combined with bioinformatics prediction analysis and q RT-PCR,it was found that hsa-mi R-382-5p and hsa-mi R-583,targeted by circRNA 001988 were upregulated, while the expression of CCDC6 which was jointly acted by hsa-mi R-382-5p and hsa-mi R-583, was downregulated by（26.20±3.92） folds and（12.61±1.48） folds in 16HBE-T and H460 cells, respectively.Conclusion CircRNA 001988 was downregulated in 16HBE-T cells transformed by benzo [a]pyrene. CircRNA 001988 may play a role as a tumor suppressor in benzo[a]pyrene-induced lung cancer and affact the expression of downstream miRNAs and mRNAs through the mechanism of endogenous competition.
出处
《环境与健康杂志》
CAS
北大核心
2016年第7期565-568,共4页
Journal of Environment and Health
基金
国家自然科学基金(81573180)
