载脂蛋白A1通过Wnt/β-catenin信号通路调控EMT在甲状腺癌转移中的作用  

ApoA1 Promotes Thyroid Carcinoma Metastasis by Modulating EMT via Wnt/β-catenin Signaling Pathway

作　　者：黄坤寨[1] 肖方森[1] 张福星[1] 张思宇[1] 张景斌[1] Huang Kunzhai;Xiao Fangsen;Zhang Fuxing(Department of General Surgery,The First Affiliated Hospital of Xiamen University Xiamen 361003,China)

机构地区：[1]厦门大学附属第一医院普外科,厦门361003

出　　处：《华中科技大学学报（医学版）》2019年第1期9-14,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：厦门市卫生计生委医学创新课题资助项目(No.2017-CXB-13)

摘　　要：目的探讨载脂蛋白A1(ApoA1)对甲状腺癌细胞侵袭和迁移的影响及可能的作用机制。方法免疫组织化学染色法检测73例甲状腺癌和50例甲状腺结节组织(对照组)ApoA1表达,实时荧光定量PCR和Western blot法检测人甲状腺乳头状癌TPC-1细胞和正常人甲状腺Nthy-ori 3-1细胞ApoA1表达;通过转染pLKO.1-shApoA1敲低TPC-1细胞ApoA1表达,Transwell小室检测细胞侵袭和迁移能力,Western blot法检测上皮细胞-间充质转化(EMT)和Wnt/β-catenin信号通路相关蛋白表达。结果甲状腺癌转移组、无转移组ApoA1染色评分明显高于对照组,转移组ApoA1染色评分又明显高于无转移组(均P<0.05)。TPC-1细胞ApoA1mRNA和蛋白表达明显高于Nthy-ori 3-1细胞,差异具有统计学意义(P<0.01)。ApoA1表达与甲状腺癌转移和临床分期有关(均P<0.05),与性别、年龄、病理分型等无关(均P>0.05)。敲低ApoA1表达可显著降低TPC-1细胞侵袭、迁移能力和抑制EMT转换能力,ApoA1-siRNA组侵袭和迁移细胞数明显少于空白对照组和空载质粒(EV)组,E-cadherin蛋白表达明显高于空白对照组和EV组,Vimentin蛋白表达明显低于空白对照组和EV组,差异均具有统计学意义(均P<0.01)。敲低ApoA1表达可明显抑制Wnt/β-catenin通路相关蛋白表达,ApoA1-siRNA组CK2α、β-catenin、Survivin蛋白表达明显低于空白对照组和EV组,差异具有统计学意义(均P<0.01)。结论抑制ApoA1表达使甲状腺癌细胞侵袭和迁移能力减弱,其作用机制可能与阻断Wnt/β-catenin信号通路、抑制细胞发生EMT有关。Objective To investigate the effects of apolipoprotein A1(ApoA1)on the invasion and migration of thyroid cancer cells and the possible mechanism.Methods The expression of ApoA1 in thyroid carcinoma(n=73)and thyroid nodule tissues(n=50 cases,control group)was assayed by immunohistochemistry.ApoA1 expression in thyroid cancer cell lines TPC-1 and normal thyroid cells Nthy-ori 3-1 was measured by qRT-PCR and Western blotting.TPC-1 cells were transfected with pLKO.1-shApoA1 to knock down the expression of ApoA1.The migratory and invasive ability of cells was measured by Transwell assays.The levels of protein related to epithelial mesenchymal transition(EMT)and Wnt/β-catenin signaling pathway were detected by Western blotting.Results ApoA1 staining scores were significantly higher in metastatic group and primary group than in the control group(all P<0.05).The mRNA and protein expression levels of ApoA1 were higher in TPC-1 cells than in the Nthy-ori 3-1 cells(all P<0.05).The expression of ApoA1 was related to the metastasis and clinical stages of thyroid carcinoma(P<0.05),but was not to age,gender and histopathology(all P >0.05).Knockdown of ApoA1 could significantly reduce the migration,invasion and EMT of TPC-1 cells.The number of migratory and invasive cells in ApoA1-siRNA group was significantly lower than that in EV and blank control group.E-cadherin protein expression was significantly higher and vimentin protein expression significantly lower in ApoA1-siRNA group than in EV and blank control group(all P<0.05).Knockdown of ApoA1 could significantly inhibit the Wnt/β-catenin signaling pathway.The expression levels of CK2α,β-catenin,survivin protein were significantly lower in ApoA1-siRNA group than in EV and blank control group(all P<0.05).Conclusion ApoA1 knockdown can inhibit the migration and invasion of thyroid carcinoma cells,and the mechanism may be related to blocking the Wnt/β-catenin signaling pathway and inhibiting the occurrence of EMT in cells.

关 键 词：载脂蛋白A1 甲状腺癌 上皮细胞-间充质转化 信号通路 

分 类 号：R736.1[医药卫生—肿瘤]