Jiangsu Provincial Key Laboratory of Geriatrics, Department of Geriatrics, the First Affiliated Hospital with Nanjing Medical University, Nanjing, China.
Department of Epidemiology, School of Public Health, Nanjing Medical University, Nanjing, China.
Cancer Med. 2019 May;8(5):2636-2645. doi: 10.1002/cam4.2073. Epub 2019 Mar 7.
To investigate the associations between the functional single nucleotide polymorphisms (SNPs) in the miR-125 family and the survival of non-small cell lung cancer (NSCLC) patients, we systematically selected six functional SNPs located in three pre-miRNAs (miR-125a, miR-125b-1, miR-125b-2). Cox proportional hazard regression analyses were conducted to estimate the crude and adjusted hazard ratios (HRs) and their 95% confidence intervals (CIs). Reporter gene luciferase assay was performed to examine the relationship between the SNPs and transcriptive activity of the miRNAs. The expression of miRNAs in different cells was detected using quantitative real-time PCR assay. We found that rs2241490 (upstream of miR-125b-1, G > A, adjusted HR = 1.24, 95%CI = 1.05-1.48, P = 0.014, in dominant model; adjusted HR = 1.18, 95%CI = 1.03-1.35, P = 0.014, in additive model), rs512932 (upstream of miR-125b-1, A > G, dominant model: adjusted HR = 1.25, 95%CI = 1.05-1.48, P = 0.013) and rs8111742 (upstream of miR-125a, G > A, dominant model: adjusted HR = 0.84, 95%CI = 0.71-1.00, P = 0.047) were associated with the prognosis of 1001 Chinese NSCLC patients. The combined analysis of the three SNPs related the number of risk alleles (rs2241490-A, rs512932-G and rs8111742-G) to death risk of NSCLC in a locus-dosage mode (P for trend <0.001). Furthermore, luciferase reporter gene assay showed significantly higher levels of luciferase activity with rs512932 variant G than that with A allele in 293T, SPC-A1 and A549 cell lines. Besides, miR-125b was highly expressed in lung cancer cells than the normal lung cell. Our study indicated that genetic variations in miR-125 family were implicated in the survival of NSCLC patients. Larger population-based and functional studies are needed to verify these findings.
为了研究 miR-125 家族中的功能性单核苷酸多态性(SNP)与非小细胞肺癌(NSCLC)患者生存之间的关系,我们系统地选择了三个前体 miRNA(miR-125a、miR-125b-1 和 miR-125b-2)中六个位于功能性 SNP。使用 Cox 比例风险回归分析来估计粗风险比(HR)和调整后的 HR(95%置信区间[CI])。进行报告基因荧光素酶测定以检查 SNP 与 miRNA 转录活性之间的关系。使用实时定量 PCR 测定检测不同细胞中 miRNA 的表达。我们发现 rs2241490(miR-125b-1 的上游,G>A,调整后的 HR=1.24,95%CI=1.05-1.48,P=0.014,在显性模型中;调整后的 HR=1.18,95%CI=1.03-1.35,P=0.014,在加性模型中),rs512932(miR-125b-1 的上游,A>G,显性模型:调整后的 HR=1.25,95%CI=1.05-1.48,P=0.013)和 rs8111742(miR-125a 的上游,G>A,显性模型:调整后的 HR=0.84,95%CI=0.71-1.00,P=0.047)与 1001 名中国 NSCLC 患者的预后相关。三个相关 SNP 的风险等位基因数(rs2241490-A、rs512932-G 和 rs8111742-G)的联合分析表明,NSCLC 的死亡风险与 NSCLC 患者的死亡风险呈剂量依赖性(P<0.001)。此外,荧光素酶报告基因测定显示,与 A 等位基因相比,rs512932 变异 G 在前 293T、SPC-A1 和 A549 细胞系中具有更高的荧光素酶活性。此外,miR-125b 在肺癌细胞中的表达水平明显高于正常肺细胞。我们的研究表明,miR-125 家族中的遗传变异与 NSCLC 患者的生存有关。需要更大的基于人群的和功能研究来验证这些发现。