甲状腺癌患者组织及手术前后血清中NR3C2和ZEB1表达水平及其与预后价值研究

Study on the Expression Level of NR3C2 and ZEB1 in Thyroid Cancer Tissue and Serum of Patients before and after Surgery and Their Prognostic Value

目的探讨核受体亚家族3C组成员2(nuclear receptor subfamily 3,group C,member 2,NR3C2)和E盒结合锌指蛋白1(E-box-bindingzincfingerprotein1,ZEB1)在甲状腺癌组织中的表达差异及手术前后血清NR3C2mRNA和ZEB1 mRNA的相对表达水平,分析其与患者预后的关系。方法选取2018年3月~2019年5月在邹城市人民医院进行手术治疗的85例甲状腺癌患者的癌组织和癌旁组织(距癌组织>3cm)样本,采用免疫组织化学法(ICC)检测癌组织和癌旁组织中NR3C2和ZEB1的表达;分别收集甲状腺癌患者(实验组)手术前、术后1周、术后1个月的血清样本,以及同期到该院体检的30例健康者(对照组)的血清样本,采用实时荧光定量PCR检测NR3C2 mRNA和ZEB1 mRNA相对表达水平;分析NR3C2mRNA和ZEB1mRNA相对表达水平与临床病理特征的关系;术前NR3C2和ZEB1表达水平与甲状腺癌患者三年生存率的关系用Kaplan-Meier法分析,采用对数秩检验进行组间生存曲线差异分析;采用多因素COX回归分析甲状腺癌患者预后影响因素。结果免疫组织化学法观察显示,癌组织中NR3C2阳性表达率(36.47%)低于癌旁组织(72.94%),癌组织中ZEB1阳性表达率(67.06%)高于癌旁组织(30.59%),差异具有统计学意义(χ^(2)=22.814,22.624,均P<0.001);实验组术前血清NR3C2 mRNA表达水平(0.55±0.14)低于对照组表达水平(0.97±0.22),术前、术后1周和术后1个月血清NR3C2mRNA表达水平(0.55±0.14,0.69±0.15,0.89±0.19)依次升高,差异具有统计学意义(t=12.038,F=95.217,P<0.001);实验组术前ZEB1 mRNA表达水平(1.88±0.28)高于对照组表达水平(1.02±0.41),术前、术后1周、术后1个月血清ZEB1mRNA表达水平(1.88±0.28,1.43±0.32,1.15±0.29)依次降低,差异具有统计学意义(t=12.716,F=130.564,P<0.001);甲状腺癌患者血清NR3C2 mRNA表达和ZEB1mRNA表达水平与TNM分期、淋巴结转移、分化程度和包膜浸润有关(t=2.449,2.081,3.938,2.212;2.013,2.348,2.029,2.096,均P<0.05);NR3C2 mRNA高表达组三年生存率(86.00%)显著高于低表达组(40.00%),ZEB1 mRNA高表达组三年生存率(35.29%)显著低于低表达组(88.24%),差异均有统计学意义(χ^(2)=4.168,5.593,均P<0.05);多因素COX回归分析显示,TNM分期、淋巴结转移、包膜浸润、NR3C2 mRNA和ZEB1 mRNA均为甲状腺癌患者预后影响因素(P<0.05)。结论甲状腺癌组织和血清中NR3C2表达下调,ZEB1表达上调,NR3C2和ZEB1表达水平影响患者预后生存率,对甲状腺癌患者预后评估具有重要意义。

Objective To investigate the expression of nuclear receptor subfamily 3,group C,member 2(NR3C2)and E-box-binding zinc finger protein 1(ZEB1)in thyroid cancer tissues,compare the relative expression of NR3C2 mRNA and ZEB1 mRNA in serum before and after surgery,and analyze their relationship with patient prognosis.Methods From March 2018 to May 2019,samples of cancer tissue and paracancerous tissue(>3cm from the cancer tissue)of 85 patients with thyroid cancer who underwent surgery in Zoucheng People’s Hospital were collected.Immunohistochemistry(ICC)was used to detect the expression of NR3C2 and ZEB1 in cancer tissue and paracancerous tissue.Serum samples were collected from thyroid cancer patients(experimental group)before surgery,1 week after surgery,and 1 month after surgery,and serum samples from 30 healthy people(control group)who went to the hospital for physical examination were also collected,real-time PCR was applied to detect the relative expression of NR3C2 mRNA and ZEB1 mRNA.The clinicopathological characteristics of patients with thyroid cancer were collected,and the relationship between the relative expression levels of NR3C2 mRNA and ZEB1 mRNA and the clinicopathological characteristics was analyzed.Kaplan-Meier method was applied to analyze the relationship between preoperative NR3C2 and ZEB1 expression levels and 3-year survival in patients with thyroid cancer,log-rank test was used to analyze the difference of survival curves between groups,and multivariate COX regression was used to analyze prognostic factors in thyroid cancer patients.Results ICC observation showed that the positive expression rate of NR3C2 in cancer tissue(36.47%)was lower than that in adjacent tissue(72.94%),and the positive expression rate of ZEB1 in cancer tissue(67.06%)was higher than that in adjacent tissue(30.59%),and the differences were statistically significant(χ^(2)=22.814,22.624,all P<0.001).The preoperative serum NR3C2 mRNA in the experimental group(0.55±0.14)was lower than that in the control group(0.97±0.22),the relative expression levels of serum NR3C2 mRNA increased in sequence before surgery,1 week after surgery,and 1 month after surgery(0.55±0.14,0.69±0.15,0.89±0.19),and the differences were statistically significant(t=12.038,F=95.217,P<0.001).The expression level of ZEB1 mRNA in the experimental group before operation(1.88±0.28)was higher than that in the control group(1.02±0.41),and the relative expression level of serum ZEB1 mRNA decreased in sequence before operation,1 week after operation,and 1 month after operation(1.88±0.28,1.43±0.32,1.15±0.29),and the differences was statistically significant(t=12.716,F=130.564,P<0.001).The relative expression of serum NR3C2 mRNA and ZEB1 RNA in patients with thyroid cancer was related to TNM stage,lymph node metastasis,degree of differentiation,and capsule invasion(t=2.449,2.081,3.938,2.212;2.013,2.348,2.029,2.096,all P<0.05).The 3-year survival rate of NR3C2 mRNA high expression group(86.00%)was obviously higher than that of low expression group(40.00%),and the 3-year survival rate of ZEB1 mRNA high expression group(35.29%)was obviously lower than that of low expression group(88.24%),the differences were statistically significant(χ^(2)=4.168,5.593,all P<0.05).Multivariate COX regression analysis showed that TNM stage,lymph node metastasis,capsular invasion,NR3C2 mRNA,and ZEB1 mRNA were all prognostic factors in patients with thyroid cancer(P<0.05).Conclusion The expression of NR3C2 in thyroid cancer tissue and serum was down-regulated,and the expression of ZEB1 was up-regulated,the expression levels of NR3C2 and ZEB1 affect the prognosis and survival rate of patients,and are of great significance for the prognosis evaluation of patients with thyroid cancer.