甲状腺乳头状癌组织中miR-338-3p、PLCD3表达及其与病理参数、上皮—间质转化、预后的关系

Expression of miR-338-3p and PLCD3 in papillary thyroid carcinoma tissues and association with pathological parameters,epithelial-mesenchymal transition and prognosis

目的探讨甲状腺乳头状癌(PTC)组织中微小核糖核酸-338-3p(miR-338-3p)、磷脂酶Cδ-3(PLCD3)表达及与病理参数、上皮—间质转化(EMT)、预后的关系。方法选取2016年1月—2020年12月商洛市中心医院乳甲外科收治接受手术切除的PTC患者152例,取其癌组织及对应癌旁组织,采用实时荧光定量聚合酶链式反应检测miR-338-3p、PLCD3 mRNA表达,免疫组织化学法检测EMT相关蛋白[N-钙黏蛋白(N-Cad)、E-钙黏蛋白(E-Cad)、波形蛋白(VIM)];分析miR-338-3p、PLCD3 mRNA表达与PTC病理参数的关系;通过Targetscan数据库预测miR-338-3p与PLCD3的结合位点,采用Pearson法和二列相关性分析miR-338-3p与PLCD3 mRNA及二者与EMT相关蛋白在PTC组织中表达的相关性;根据PTC组织miR-338-3p、PLCD3 mRNA表达均值分为高/低miR-338-3p、PLCD3 mRNA表达组,通过Kaplan-Meier法绘制高/低miR-338-3p、PLCD3 mRNA表达PTC患者无病生存率(DFS)生存曲线;通过Cox回归分析PTC患者DFS的影响因素。结果与癌旁组织比较,PTC组织miR-338-3p和E-Cad蛋白阳性表达率降低,PLCD3 mRNA和N-Cad、VIM蛋白阳性表达率升高(t/χ^(2)/P=32.875/<0.001、15.575/<0.001、37.351/<0.001、21.984/<0.001、16.604/<0.001);miR-338-3p与PLCD3 mRNA在PTC组织中表达呈负相关(r/P=-0.712/<0.001)。PTC组织中miR-338-3p与N-Cad、VIM蛋白阳性表达呈负相关,与E-Cad蛋白阳性表达呈正相关(r/P=-0.642/<0.001、-0.617/<0.001、0.622/<0.001);PTC组织中PLCD3 mRNA与N-Cad、VIM蛋白阳性表达呈正相关,与E-Cad蛋白阳性表达呈负相关(r/P=0.657/<0.001、0.624/<0.001、-0.632/<0.001)。不同TNM分期、淋巴结转移的PTC组织miR-338-3p、PLCD3 mRNA表达差异有统计学意义(F/t/P=30.778/<0.001、3.430/0.001)。miR-338-3p高表达组3年DFS高于低表达组,PLCD3 mRNA高表达组3年DFS低于低表达组(χ^(2)/P=15.615/<0.001、16.088/<0.001)。TNM分期Ⅲ期、淋巴结转移、PLCD3 mRNA≥1.83为PTC患者DFS的独立危险因素[HR(95%CI)=3.127(1.361~7.604)、2.546(1.115~5.817)、2.854(1.178~6.919)],miR-338-3p≥0.57为独立保护因素[HR(95%CI)=0.306(0.116~0.804)]。结论PTC组织中miR-338-3p低表达和PLCD3 mRNA高表达,与TNM分期、淋巴结转移、EMT和预后有关,可能成为PTC诊治新靶点。

Objective To investigate the expression of micro ribonucleic acid-338-3p(miR-338-3p)and phospholipase C delta-3(PLCD3)in papillary thyroid carcinoma(PTC)tissues and their relationship with pathological parameters,epithelial-mesenchymal transition(EMT)and prognosis.Methods 152 cases of PTC patients admitted to the Breast and Thyroid Surgery Department of Shangluo Central Hospital who underwent surgical resection from January 2016 to December 2020 were selected,and real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of miR-338-3p and PLCD3 mRNA in cancer tissues and corresponding para-cancerous tissues,and immunohistochemistry was used to detect the expression of EMT-associated proteins[N-calmodulin(N-Cad),E-calmodulin(E-Cad),and vimentin(VIM)].The relationship between miR-338-3p,PLCD3 mRNA expression and pathological parameters of PTC were analyzed.The binding sites of miR-338-3p and PLCD3 were predicted by Targetscan database,and the correlation between miR-338-3p and PLCD3 mRNA and the expression of both with EMT-related proteins in PTC tissues was analyzed by Pearson's method and dichotomous correlation.PTC tissues were divided into high/low miR-338-3p,PLCD3 mRNA expression groups according to the mean values of miR-338-3p,PLCD3 mRNA expression,and the survival curves of disease-free survival(DFS)of PTC patients with high/low miR-338-3p,PLCD3 mRNA expression were plotted by Kaplan-Meier method.Factors of DFS in PTC patients were analyzed by Cox regression.Results Compared with para-cancerous tissues,miR-338-3p expression and E-Cad protein positive expression rate were decreased in PTC tissues,and PLCD3 mRNA expression and N-Cad and VIM protein positive expression rate were increased(t/χ^(2)/P=32.875/<0.001,15.575/<0.001,37.351/<0.001,21.984/<0.001,16.604/<0.001).miR-338-3p was negatively correlated with PLCD3 mRNA expression in PTC tissues(r/P=-0.712/<0.001).miR-338-3p was negatively correlated with the positive expression rate of N-Cad and VIM proteins,and positively correlated with that of E-Cad proteins in PTC tissues(r/P=-0.642/<0.001,-0.617/<0.001,0.622/<0.001);PLCD3 mRNA was negatively correlated with N-Cad in PTC tissues,VIM protein positive expression rate was positively correlated with E-Cad protein positive expression rate(r/P=0.657/<0.001,0.624/<0.001,-0.632/<0.001).Comparison of miR-338-3p and PLCD3 mRNA expression in PTC tissues with different TNM stages and lymph node metastasis showed differences(F/t/P=30.778/<0.001,3.430/0.001).3-year DFS was higher in the miR-338-3p high-expression group than that in the miR-338-3p low-expression group,and 3-year DFS was lower in the PLCD3 mRNA high-expression group than that in the PLCD3 mRNA low-expression group(χ^(2)/P=15.615/<0.001,16.088/<0.001).TNM stageⅢ,lymph node metastasis,and PLCD3 mRNA≥1.83 were independent risk factors for DFS in PTC patients,and miR-338-3p≥0.57 was an independent protective factor[HR(95%CI)=3.127(1.361-7.604),2.546(1.115-5.817),2.854(1.178-6.919),0.306(0.116-0.804)].Conclusion Low expression of miR-338-3p and high expression of PLCD3 mRNA in PTC tissues are associated with TNM staging,lymph node metastasis,EMT and prognosis,and may become a new target for PTC diagnosis and treatment.