NRG1基因多态性协同miRNA-142水平与甲状腺癌侵袭转移的关系研究

Relationship of NRG1 Gene Polymorphism and miRNA-142 Expression with Invasion and Metastasis of Thyroid Cancer

[目的]分析神经调节素1(NRG1)基因多态性协同微小核糖核酸miRNA-142(miR-142)水平与甲状腺癌侵袭转移的关系。[方法]选择120例甲状腺癌患者作为病例组,另选同期体检的120名健康者作为对照组。采用聚合酶链反应-限制性片段长度多态性法(PCR-RFLP)测定NRG1基因3个不同位点的单核苷酸多态性(SNP),TaqMan等位基因分型技术分析基因分型及基因频率,实时定量PCR法测定血清miR-142水平。对比各组间NRG1基因多态性、miR-142水平差异,进行Logistic回归交互作用分析,绘制受试者工作特征(ROC)曲线,分析NRG1基因多态性协同miR-142水平预测甲状腺癌患者包膜浸润、淋巴结转移的价值。[结果]病例组NRG1基因rs6994992、rs7835688位点基因型分布频率与对照组比较,差异无统计学意义(P>0.05);病例组NRG1基因rs3924999位点携带A等位基因突变的基因型AA+GA与野生型GG相比,差异有统计学意义(P<0.05),携带AA+GA基因型的发病风险增加2.664倍(95%CI:1.062~6.684);病例组NRG1基因rs3924999位点等位基因频率分布与对照组比较,差异有统计学意义(P<0.05),等位基因A突变可使发病风险增加2.056倍(95%CI:1.388~3.045)。病例组血清miR-142表达水平(0.61±0.19)低于对照组(1.00±0.21)(P<0.05)。包膜浸润组、淋巴结转移组患者NRG1基因rs3924999位点基因频率分布分别与无包膜浸润组、无淋巴结转移组相比,差异均有统计学意义(P<0.05)。Logistic回归交互分析证实NRG1基因rs3924999位点多态性、miR-142之间存在交互作用,NRG1基因rs3924999位点多态性协同miR-142水平预测甲状腺癌患者包膜浸润、淋巴结转移的AUC分别为0.785(95%CI:0.701~0.869)、0.797(95%CI:0.715~0.897)。[结论]NRG1基因rs3924999位点多态性、miR-142水平与甲状腺癌发生、侵袭、转移密切相关,两者存在交互作用,且协同有助于预测甲状腺患者包膜浸润、淋巴结转移。

[Objective]To analyze the relationship of neurotonin 1(NRG1)gene polymorphism and miRNA-142(miR-142)expression with the invasion and metastasis of thyroid cancer.[Methods]One hundred and twenty patients with thyroid cancer(case group)and 120 healthy subjects(control group)were enrolled in the study.Single nucleotide polymorphisms(SNP)at 3 different sites of NRG1 gene was detected by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP),genotyping and gene frequency was analyzed by Taq Man allele typing technology,serum miR-142 level was determined by real-time quantitative RT-PCR.The NRG1 gene polymorphism and miR-142 expression level between two groups were compared,and the interaction analysis of Logistic regression was performed.The receiver operating characteristic(ROC)curve was used to analyze the value of NRG1 gene polymorphism and miR-142 level in predicting capsule invasion and lymph node metastasis in patients with thyroid cancer.[Results]There was no significant difference in the genotype frequencies of rs6994992 and rs7835688 of NRG1 gene between the case group and the control group(P>0.05).There was significant difference in the frequency of AA+GA in rs3924999 locus of NRG1 gene between the case group and control group(P<0.05),and the risk of thyroid cancer for those carrying AA+GA genotype increased by 2.664 times(95%CI:1.062~6.684).The frequency distribution of NRG1 gene rs3924999 was statistically significant(P<0.05)between the case group and control group;and the mutation of allele A increased the risk of disease by 2.056 times(95%CI:1.388-3.045).The level of serum miR-142 expression in the case group(0.61±0.19)was lower than that in the control group(1.00±0.21)(P<0.05).The frequency distribution of rs3924999 of NRG1 gene in patients with capsular infiltration and lymph node metastasis was significantly different from that in patients without capsular infiltration and lymph node metastasis(P<0.05).Logistic regression analysis showed that there was interaction between rs3924999 polymorphism of NRG1 gene and miR-142.The area under the ROC curve(AUC)of rs3924999 polymorphism of NRG1 gene and miR-142 level in predicting capsule invasion and lymph node metastasis of thyroid cancer patients was 0.785(95%CI:0.701~0.869)and 0.797(95%CI:0.715~0.897),respectively.[Conclusion]The rs3924999 polymorphism of NRG1 gene and the expression level of miR-142 are closely related to the occurrence,invasion and metastasis of thyroid cancer.The detection of NRG1 gene rs3924999 polymorphism and miR-142 expression level may be helpful to predict capsule infiltration and lymph node metastasis in thyroid cancer patients.