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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 study the expression of circRNA 001988 and its downstream target genes in 16HBE-T cells from malignant transformation of benzo [a] pyrene cells and to explore the possible role of this circRNA in carcinogenesis. Methods The expression of circRNA 001988 in 16HBE-T was detected by q RT-PCR. The expression of circRNA 001988 was detected in human lung cancer cell lines A549, H460 and H446. Meanwhile, the expression of circRNA 001988 was detected in 20 lung cancer samples. Bioinformatics analysis software mi RSystem and RegRNA were used to predict the target microRNAs and their interaction with circRNA 001988. The qRT-PCR was used to detect the expression of microRNA and mRNA in 16HBE-T and H460. Results Compared with 16HBE, the expression of circRNA001988 in 16HBE-T was down-regulated by (6.54 ± 0.75) fold, and the difference was statistically significant (P <0.01), while downregulated in A549 and H460 cells (2.06 ± 0.38) Times, (3.02 ± 0.51) times, the differences were statistically significant (P <0.05). The expression of circRNA001988 in lung cancer was decreased by an average of (2.11 ± 0.71) fold compared with that of the adjacent non-cancerous tissues (P <0.05). Combined with predictive bioinformatics analysis and qRT-PCR detection, the expression of hsa-mi R-382-5p and hsa-miR-583 targeted by 16S rRNA was up-regulated in 16HBE-T and H460 cells compared with 16HBE The mRNA expression of CCDC6 with hsa-miR-382-5p and hsa-miR-583 decreased by (26.20 ± 3.92) and (12.61 ± 1.48) fold, respectively. CONCLUSION: The expression of circRNA 001988 in 16HBE-T cells with malignant transformation of benzo [a] pyrene is significantly reduced. It may play a role of tumor suppressor in lung carcinogenesis induced by benzo [a] pyrene, The mechanism of competition affects the expression of microRNA and mRNA downstream of it.