乳腺癌组织NKD1与β-catenin表达临床意义
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摘要
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 drosophila1(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,and 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 ofβ-catenin and EMT-related proteins in MCF-7 cells.The relationship between NKD1 andβ-catenin in IBC 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(χ~2=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)and E-cadherin(t=10.127,P<0.001)in NKD1 activation group were higher than those of no-load group;the expression of Wnt1(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
OBJECTIVE Naked cuticle drosophila1(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,and 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 ofβ-catenin and EMT-related proteins in MCF-7 cells.The relationship between NKD1 andβ-catenin in IBC 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(χ~2=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)and E-cadherin(t=10.127,P<0.001)in NKD1 activation group were higher than those of no-load group;the expression of Wnt1(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
引文
[1]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
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[11]Clevers H,Nusse R.Wnt/β-catenin signaling and disease[J].Cell,2012,149(6):1192-1205.
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[13]张俊霞,徐金升,王静,等.Wnt通路活化对肾透明细胞癌786-o细胞体外增殖与转移潜能影响及其机制探讨[J].中华肿瘤防治杂志,2016,23(3):159-163.
[14]Akalay I,Tan TZ,Kumar P,et al.Targeting WNT1-inducible signaling pathway protein 2alters human breast cancer cell susceptibility to specific lysis through regulation of KLF-4and miR-7expression[J].Oncogene,2015,34(17):2261-2271.
[15]唐凤娟,许三鹏,王国平,等.Wnt5a和Wnt11在肺癌中的表达及意义[J].临床与实验病理学杂志,2017,33(10):1082-1086.
[16]Akter H,Park M,Kwon OS,et al.Activation of matrix metalloproteinase-9(MMP-9)by neurotensin promotes cell invasion and migration through ERK pathway in gastric cancer[J].Tumor Biol,2015,36(8):6053-6062.
[17]Lee J,Hahm ER,Marcus AI,et al.Withaferin A inhibits experimental epithelial-mesenchymal transition in MCF-10Acells and suppresses vimentin protein level in vivo in breast tumors[J].Mol Carcinogen,2015,54(6):417-429.
[18]Ye Z,Zhang X,Luo Y,et al.Prognostic values of vimentin expression and Its clinicopathological significance in non-small cell lung cancer:a Meta-analysis of observational studies with 4 118cases[J].Plos One,2016,11(9):e0163162.
[19]Coopman P,Djiane A.Adherens junction and E-Cadherin complex regulation by epithelial polarity[J].J Marine Syst,2016,73(18):3535-3553.
[2]Ferlay J,Soerjomataram I,Dikshit R,et al.Cancer incidence and mortality worldwide:sources,methods and major patterns in GLOBOCAN 2012[J].Int J Cancer,2015,136(5):E359-E386.
[3]Larraguibel J,Weiss AR,Pasula DJ,et al.Wnt ligand-dependent activation of the negative feedback regulator Nkd1[J].Mol Bio Cell,2015,26(12):2375-2384.
[4]何艳姣,徐妙生,刘朝霞,等.β-catenin基因在乳腺癌中的表达和突变[J].肿瘤研究与临床,2011,23(2):97-99.
[5]饶翔,杨丞.鼻咽癌组织HSP27与β-catenin及Vimentin表达相关性研究[J].中华肿瘤防治杂志,2017,24(3):183-187.
[6]林山,蒋晓芳,王亚萍,等.NKD1和β-catenin蛋白在结直肠癌组织中的表达及意义[J].山东医药,2014,54(25):38-40.
[7]Maruyama K,Ochiai A,Akimoto S,et al.Cytoplasmic beta-catenin accumulation as a predictor of hematogenous metastasis in human colorectal cancer[J].Oncology,2000,59(4):302-309.
[8]Wharton KAJr,Zimmermann G,Rousset R,et al.Vertebrate proteins related to drosophila naked cuticle bind dishevelled and antagonize Wnt signaling[J].Dev Biol,2001,234(1):93-106.
[9]Guo M,Zhang X,Wang G,et al.miR-603promotes glioma cell growth via Wnt/β-catenin pathway by inhibiting WIF1and CT-NNBIP1[J].Cancer Lett,2015,360(1):76-86.
[10]Sugioka K,Mizumoto K,Sawa H.Wnt regulates spindle asymmetry to generate asymmetric nuclearβ-catenin in C.elegans[J].Cell,2011,146(6):942-954.
[11]Clevers H,Nusse R.Wnt/β-catenin signaling and disease[J].Cell,2012,149(6):1192-1205.
[12]Larraguibel J,Weiss AR,Pasula DJ,et al.Wnt ligand-dependent activation of the negative feedback regulator Nkd1[J].Mol Bio Cell,2015,26(12):2375-2384.
[13]张俊霞,徐金升,王静,等.Wnt通路活化对肾透明细胞癌786-o细胞体外增殖与转移潜能影响及其机制探讨[J].中华肿瘤防治杂志,2016,23(3):159-163.
[14]Akalay I,Tan TZ,Kumar P,et al.Targeting WNT1-inducible signaling pathway protein 2alters human breast cancer cell susceptibility to specific lysis through regulation of KLF-4and miR-7expression[J].Oncogene,2015,34(17):2261-2271.
[15]唐凤娟,许三鹏,王国平,等.Wnt5a和Wnt11在肺癌中的表达及意义[J].临床与实验病理学杂志,2017,33(10):1082-1086.
[16]Akter H,Park M,Kwon OS,et al.Activation of matrix metalloproteinase-9(MMP-9)by neurotensin promotes cell invasion and migration through ERK pathway in gastric cancer[J].Tumor Biol,2015,36(8):6053-6062.
[17]Lee J,Hahm ER,Marcus AI,et al.Withaferin A inhibits experimental epithelial-mesenchymal transition in MCF-10Acells and suppresses vimentin protein level in vivo in breast tumors[J].Mol Carcinogen,2015,54(6):417-429.
[18]Ye Z,Zhang X,Luo Y,et al.Prognostic values of vimentin expression and Its clinicopathological significance in non-small cell lung cancer:a Meta-analysis of observational studies with 4 118cases[J].Plos One,2016,11(9):e0163162.
[19]Coopman P,Djiane A.Adherens junction and E-Cadherin complex regulation by epithelial polarity[J].J Marine Syst,2016,73(18):3535-3553.