乳腺癌组织NKD1与β-catenin表达临床意义

NKD1 and β-catenin expression in the breast cancer tissue and its significance

目的裸角质膜同源蛋白(naked cuticle drosophila1,NKD1)是Wnt/β-catenin信号通路上重要的负向调控因子之一,β-catenin在乳腺癌组织中存在异常表达,且影响乳腺癌细胞生物学特征。本研究探讨NKD1与β-catenin在乳腺癌组织中表达及意义。方法收集2014-01-01-2015-12-30沧州市中心医院手术切除的200例乳腺癌肿瘤组织标本,其中浸润性乳腺癌(invasive breast cancer,IBC)150例、导管内癌50例,同时选取癌旁正常乳腺组织标本50例作为对照,采用免疫组化法、蛋白质印迹法和聚合酶链反应法检测所有组织中NKD1与β-catenin表达,蛋白质印迹法检测NKD1表达对MCF-7细胞中β-catenin和上皮间质转化(epithelial mesenchymal transformation,EMT)相关蛋白表达影响,分析IBC组织中NKD1与β-catenin关系及与患者临床病理参数关系。结果 IBC组织NKD1蛋白(t=6.237,P=0.025)和NKD1mRNA(t=9.428,P<0.001)相对表达量低于正常乳腺组织;IBC组织β-catenin蛋白表达量(t=5.208,P=0.011)高于正常乳腺组织。IBC和导管内癌NKD1阳性细胞比例低于正常乳腺组织,F=13.142,P<0.001;IBC和导管内癌β-catenin异位表达阳性细胞比例高于正常乳腺组织,F=9.034,P=0.011。IBC组织NKD1表达量与β-catenin异位表达量呈负相关(r=-0.928,P=0.011)。NKD1高低表达与IBC患者肿瘤分化程度(χ2=7.128,P=0.006)、淋巴结转移(χ2=7.770,P=0.005)和TNM分期(χ2=6.489,P=0.011)有关联;β-catenin异位高低表达与IBC患者肿瘤分化程度(χ2=13.264,P<0.001)、淋巴结转移(χ2=15.748,P<0.001)和TNM分期(χ2=5.725,P=0.015)有关联。NKD1激活组NKD1(t=13.264,P<0.001)和E-cadherin(t=10.127,P<0.001)蛋白表达高于空载组;NKD1激活组Wnt1(t=15.324,P<0.001)、MMP-9(t=16.102,P<0.001)和vimentin(t=10.946,P<0.001)蛋白表达低于空载组。结论 NKD1在浸润性乳腺癌组织中呈低表达,β-catenin在浸润性乳腺癌组织中高表达,其机制可能与Wnt信号通路有关。

OBJECTIVE Naked cuticle drosophilal(NKD1)is one of the important negative regulators of Wnt/β-catenin signaling pathway,β-catenin is abnormally expressed in breast cancer tissues and affects breast cancer cells.In this paper,we studied the expression of NKD1 andβ-catenin in the breast cancer tissue,a nd to explore the correlation and significance.METHODS Totally 200 cases of breast cancer tumor tissue were collected from the Central Hospital of Cangzhou City from 1 January 2014 to 30 December 2015,including 150 cases of invasive breast cancer(IBC)and 50 cases of intraductal carcinoma.At the same time,50 specimens of normal breast tissue adjacent to the cancer were selected as controls.Immunohistochemistry,Western blotting and polymerase chain reaction were used to detect the expression of NKD1 andβ-catenin in all tissues.Western blotting was used to detect the effect of NKD1 expression on the expression of β-catenin and EMT-related proteins in MCF-7 cells.The relationship between NKD1 andβ-catenin in IEC tissues and its relationship with clinicopathological parameters were analyzed.RESULTS The relative expression of NKD1 protein(t=6.237,P=0.025)and NKD1 mRNA(t=9.428,P<0.001)in IBC tissues were lower than those in normal breast tissues,while the expression ofβ-catenin protein in IBC tissues(t=5.208,P=0.011)was higher than that in normal breast tissues.The proportion of NKD1 positive cells in IBC and intraductal cancer was lower than that in normal breast tissue.F=13.142,P<0.001;the proportion ofβ-catenin positive cells in IBC and intraductal cancer was higher than that in normal breast tissue,F=9.034,P<0.001.The expression of NKD1 in IBC tissues was negatively correlated with the ectopic expression ofβ-catenin(r=0.928,P=0.011).The expression of NKD1 was related with the degree of differentiation(χ^2=7.128,P=0.006),lymph node metastasis(χ^2=7.770,P=0.005)and TNM stage(χ^2=6.489,P=0.011)of IBC patients;heterotopic expression ofβ-catenin was associated with the degree of differentiation(t=13.264,P<0.001),lymph node metastasis(χ^2=15.748,P<0.001)and TNM stage(χ^2=5.725,P=0.015)in IBC patients.The expression of NKD1(t=13.264,P<0.001)andβ-cadherin(t=10.127,P<0.001)in NKD1 activation group were higher than those of no-load group;the expression of Wntl(t=15.324,P<0.001),MMP-9(t=16.102,P<0.001)and vimentin(t=10.946,P<0.001)in NKD1 activation group were lower than those of no-load group.CONCLUSIONS NKD1 is low in breast cancer tissues,butβ-catenin is higher.The intrinsic mechanism may be related to the Wnt signaling pathway.