摘要
目的探讨结直肠癌组织中膜联蛋白A10(ANXA10)与肿瘤生长转移的关系。方法选取唐山市工人医院肿瘤外科在2018年1月至2019年10月手术切除的结直肠腺癌标本进行研究,患者均为初诊患者。75例结直肠癌患者中男45例,女30例,年龄(57.69±8.32)岁。免疫组织化学技术(IHC)检测75例结直肠癌肿瘤及癌旁正常肠黏膜石蜡组织中ANXA10蛋白表达,分析ANXA10与患者临床病理特征的关系。应用全基因克隆技术上调人结直肠癌细胞株SW480中ANXA10的水平;新型四唑化合物(MTS)法检测细胞活性,划痕实验及Transwell小室实验检测肿瘤细胞迁移及侵袭能力。实时定量聚合酶链反应(Real-time PCR)及蛋白质印迹法(Western blot)技术检测肿瘤细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)、N-钙黏素(N-cadherin)、E-钙黏素(E-cadherin)的表达。应用SPSS 21.0软件对数据采用t检验、单因素方差分析。结果结直肠癌组织中ANXA10蛋白阳性率(22.67%)低于癌旁正常肠黏膜(54.67%,χ^2=16.190,P<0.01);ANXA10表达与患者的肿瘤分化(高中分化36.36%,低未分化11.90%)、淋巴结转移(阳性5.13%,阴性41.67%)及临床分期(Ⅰ~Ⅱ期58.33%,Ⅲ~Ⅳ期5.88%)明显相关(χ^2=6.307,P<0.05,χ^2=14.258,P<0.01,χ^2=25.614,P<0.01)。ANXA10在人结直肠癌细胞株中低于正常肠黏膜细胞株HIEC,SW480细胞中ANXA10表达最低(F=31.714、85.211,P<0.01)。转染后SW480细胞中ANXA10表达明显增高(F=1774.586、1559.994,P<0.01);转染48 h后转染组SW480细胞的活性明显低于对照组(F=271.017、215.699,P<0.01);转染组SW480细胞的迁移能力、侵袭能力明显低于对照组(F=1204.876、70.137,P<0.01)。转染组SW480细胞中PCNA、N-cadherin、MMP-2的mRNA和蛋白水平均明显低于对照组(mRNA:F=35.974,P<0.01;F=8.933,P<0.05;F=21.868,P<0.01;蛋白:F=100.484,P<0.01;F=90.413,P<0.01;F=7.990,P<0.05);而E-cadherin的mRNA和蛋白水平在转染组明显高于对照组(mRNA:F=40.375,P<0.01;蛋白:F=128.487,
Objective To investigate the relationship between annexin A10(ANXA10)and tumor growth and metastasis in colorectal cancer.Methods Among 75 patients with colorectal cancer,45 were male and 30 were female,and the age was(57.69±8.32)years.Immunohistochemical technique(IHC)was used to detect the expression of ANXA10 protein in 75 cases of colorectal cancer tumors and adjacent normal intestinal mucosa paraffin tissue,and the relationship between ANXA10 and clinicopathological characteristics of patients was analyzed.The whole gene cloning technology was used to increase the level of ANXA10 in human colorectal cancer cell line SW480;3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)was used to detect cell viability,scratch test and Transwell test were used to detect tumor cell migration and invasion ability.Real-time quantitative polymerase chain reaction(Real-time PCR)and Western blotting were used to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),N-cadherin,and E-cadherin.SPSS 21.0 software was used to test the data with t-test and one-way ANOVA.P<0.05 was considered as statistically significant difference.Results The positive rate of ANXA10 protein in colorectal cancer tissues(22.67%)was lower than that in normal intestinal mucosa adjacent to the cancer(54.67%,χ^2=16.190,P<0.01);the expression of ANXA10 was related to tumor differentiation(Well differentiated 36.36%,Poor differentiated 11.90%),lymph node metastasis(Positive 5.13%,Negative 41.67%),and clinical stage in patients(Ⅰ-Ⅱphase 58.33%,Ⅲ-Ⅳphase 5.88%,χ^2=6.307,P<0.05,χ^2=14.258,P<0.01,χ^2=25.614,P<0.01).The expression of ANXA10 was lower in human colorectal cancer cell lines than in normal intestinal mucosa cell line HIEC,and ANXA10 expression was lowest in SW480 cells(F=31.714,P<0.01;F=85.211,P<0.01).The expression of ANXA10 in SW480 cells significantly increased after transfection(F=1774.586,P<0.01;F=1559.994,P<0.01);the activity of SW480 cells in th
作者
马鹏程
王爱军
吴肖
张超
张鸣
Ma Pengcheng;Wang Aijun;Wu Xiao;Zhang Chao;Zhang Ming(Department of Oncology Surgery,Tangshan Gongren Hospital,Tangshan 063000,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第2期232-235,共4页
Chinese Journal of Experimental Surgery
关键词
结直肠癌
膜联蛋白A10
增殖
转移
临床病理特征
Colorectal cancer
Annexin A10
Proliferation
Metastasis
Clinicopathological features
