非小细胞肺癌组织中微小RNA-423-3p的拷贝数变异和rs6505162位点多态性分析

Analysis of copy number variation and rs6505162 polymorphism of microRNA-423-3p in non-small cell lung cancer tissues

目的探讨非小细胞肺癌(NSCLC)组织中微小RNA-423-3p(miR-423-3p)的拷贝数变异(CNV)和rs6505162位点单核苷酸多态性(SNP)。方法采用cBioPortal在线分析癌症基因组图谱TCGA数据库中NSCLC组织miR-423-3p的CNV表现及与临床病理特征的关系;收集本院2017年1月至2018年12月经病理确诊的146例NSCLC患者(NSCLC组)和151例健康体检者(对照组)的外周血标本,采用聚合酶链式反应限制性片段长度多态性(PCR-RFLP)检测rs6505162位点SNP的基因分型并进行Hardy-Weinberg平衡检验,分析rs6505162等位基因A在不同遗传模型[等位基因模型(A vs. C)、纯合子模型(AA vs. CC)、杂合子模型(CA vs. CC)、显性遗传模型(CA+AA vs. CC)和隐性遗传模型(AA vs. CC+CA)]下与NSCLC易感性的关系。结果经cBioPortal在线分析发现1144例组织的miR-423-3p CNV表现:缺失209例(Deep Deletion 1例+Shallow Deletion 208例)、正常(Diploid)543例和扩增392例(Gain 384例+Amplification 8例)。NSCLC组织中CNV分布与病理类型、性别和N分期有关(P<0.05),而与TNM分期、T分期、M分期和生存状况无关(P>0.05)。两组rs6505162基因型的分布符合Hardy-Weinberg平衡。NSCLC组携带基因型CA、AA的频率高于对照组(56.16%、17.81%vs. 49.67%、11.26%,P<0.05),携带等位基因A的频率高于对照组(45.89%vs. 36.09%,P<0.05)。rs6505162位点的等位基因A在隐性遗传模型下与NSCLC的易感性无关(P>0.05),而在等位基因模型(OR=1.502, 95%CI:1.081~2.087)、纯合子模型(OR=2.375, 95%CI:1.139~4.952)、杂合子模型(OR=1.698, 95%CI:1.015~2.839)和显性遗传模型(OR=1.823, 95%CI:1.113~2.986)下可升高NSCLC的发病风险。结论 miR-423-3p CNV可能参与了NSCLC的发生发展且其rs6505162与NSCLC易感性有关,其中携带突变等位基因A的NSCLC发生风险升高,在肺癌易感人群筛查中有一定价值。

Objective To investigate the copy number variation(CNV) and single nucleotide polymorphism(SNP) rs6505162 of microRNA-423-3 p(miR-423-3 p) in non-small cell lung cancer(NSCLC) tissues. Methods On-line analysis by cBioPortal was applied to reveal CNV manifestations of miR-423-3 p in NSCLC tissues from The Cancer Genome Atlas(TCGA) database and their relationship with clinicopathological features. Peripheral blood samples of 146 NSCLC patients confirmed by pathology(NSCLC group) and 151 healthy physical examinees(Control group) from January 2017 to December 2018 were collected. The genotype of rs6505162 at the polymorphic site of miR-423-3 p was detected by polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP). Hardy-Weinberg equilibrium was used to analyze the genetic balance of this SNP. The relationship between rs6505162 allele A and NSCLC susceptibility was analyzed under different genetic models including allele model(A vs. C), homozygote model(AA vs. CC), heterozygote model(CA vs. CC), dominant genetic model(CA+AA vs. CC) and recessive genetic model(AA vs. CC+CA). Results CNV manifestations of miR-423-3 p in 1144 tissues were found by cBioPortal on-line analysis as follows: 209 cases of deletion(Deep Deletion 1 case+Shallow Deletion 208 cases), 543 cases of normal(Diploid) and 392 cases of amplification(Gain 384 cases+Amplification 8 cases). The distribution of CNV in NSCLC was related to pathological type, gender and N stage(P<0.05), but not to TNM stage, T stage, M stage and survival status(P>0.05). The distribution of miR-423-3 p rs6505162 genotypes in both groups conformed to Hardy-Weinberg equilibrium. The frequencies of CA and AA in NSCLC group were higher than those in Control group(56.16%, 17.81% vs. 49.67%, 11.26%, P<0.05), and the frequency of allele A in NSCLC group were higher than that in Control group(45.89% vs. 36.09%, P<0.05). Allele A at the site of miR-423-3 p rs6505162 was not associated with NSCLC susceptibility in recessive genetic model but in allele model(OR=1.502, 95% CI: 1.081-2.087), homozygote model(OR=2.375, 95%CI: 11.139-4.952), heterozozygote model(OR=1.698, 95%CI: 1.015-2.839) and dominant genetic model(OR=1.823, 95% CI: 1.113~2.986) increased the risk of NSCLC. Conclusion MiR-423-3 p CNV may be involved in the occurrence and development of NSCLC, and rs6505162 is associated with the susceptibility of NSCLC. The risk of NSCLC carrying mutant alleles is increased, which has a certain value in screening lung cancer susceptible population.