芳基烃受体核转位因子样蛋白2在子宫内膜癌中的表达

doi:10.6040/j.issn.1671-7554.0.2021.1475

摘要/Abstract

摘要： 目的探讨芳基烃受体核转位因子样蛋白2(ARNTL2)在子宫内膜癌中的表达、作用及其可能的信号通路。方法采用免疫组织化学SP法、RT-PCR和Western blotting实验检测子宫内膜癌及正常内膜组织中ARNTL2的表达;小干扰RNA(Si-ARNTL2)敲低ARNTL2 的表达,CCK-8实验检测细胞增殖能力,划痕愈伤实验和小室穿透实验(Transwell)检测细胞迁移和侵袭能力;Western blotting检测细胞上皮间质转化指标:N-钙黏蛋白(N-cad)、E-钙黏蛋白(E-cad)、Vimentin蛋白、Snail蛋白、Slug蛋白及Wnt/β-catenin信号通路相关蛋白(Wnt蛋白、β-连环蛋白)的表达。结果免疫组织化学、RT-PCR和Western blotting结果均显示,ARNTL2在子宫内膜癌组织中表达上调(t免疫组织化学=2.213, P=0.033;t_RT-PCR=4.766, P<0.001;t_{Western blotting}=2.659,P=0.012);Si-ARNTL2转染可成功降低子宫内膜癌细胞系(KLE和Ishikawa)ARNTL2的表达。敲低ARNTL2后,CCK8实验结果显示,两种细胞的吸光度值下降(P均<0.05);划痕愈伤实验结果显示,两种细胞的划痕愈合率下降(P均<0.001);Transwell侵袭及迁移实验结果均显示,两种细胞穿膜细胞数减少(P均<0.01);Western blotting实验结果显示,两细胞系E-cad表达上升(P均<0.05),N-cad、Vimentin、Snail、Slug蛋白表达下降(P均<0.05),两细胞Wnt、β-catenin蛋白的表达均下降(P均<0.05)。结论 ARNTL2在子宫内膜癌组织中的表达升高,降低子宫内膜癌细胞中ARNTL2的表达可抑制子宫内膜癌的增殖和转移,其可能通过Wnt/β-catenin通路发挥作用。

关键词: 芳基烃受体核转位因子样蛋白2, 子宫内膜癌, 增殖, 转移, 上皮间质转化, 信号通路

Abstract: Objective To explore the expression and function of aryl hydrocarbon receptor nuclear translocator-like 2(ARNTL2)in endometrial cancer(EC)and its possible signal pathway. Methods The expression of ARNTL2 in EC and normal tissues was detected with immunohistochemistry, RT-PCR and Western blotting. After ARNTL2 knock down with Si-RNA(Si-ARNTL2), cell proliferation, invasion and migration were detected with CCK-8 assay, scratch healing test and Transwell assay, respectively. The epithelial-mesenchymal transition(EMT)indicators, including E-cad, N-cad, Vimentin, Snail and Slug, and Wnt/β-catenin signal pathway related proteins, including Wnt and β-catenin, were detected with Western blotting. Results Immunohistochemistry, RT-PCR and Western blotting showed that the expression of ARNTL2 was up-regulated in EC tissue(t_{Immunohistochemistry}=2.213, P=0.033; t_RT-PCR=4.766, P<0.001; t_{Western blotting}=2.659, P=0.012). Si-ARNTL2 transfection successfully reduced ARNTL2 expression in EC cell lines(KLE and Ishikawa). After knockdown of ARNTL2, CCK8 assay showed reduction in the absorbance of both cell lines(P<0.05); Scratch healing test showed a suppression on wound healing rate(P<0.001); Transwell invasion and migration assay showed that the number of transmembrane cells decreased(P<0.01). Western blotting showed that the expression of E-cad in the two cell lines increased, while the expressions of N-cad, Vimentin, Snail and Slug decreased(P<0.05), and the expressions of Wnt and β-catenin decreased(P<0.05). Conclusion The expression of ARNTL2 is higher in EC tissue than in normal tissue. Knockdown of ARNTL2 can inhibit cell proliferation and metastasis in EC. ARNTL2 may promote tumor proliferation and metastasis through Wnt/β-catenin pathway.

Key words: Aryl hydrocarbon receptor nuclear translocator-like 2, Endometrial cancer, Proliferation, Metastasis, Epithelial-mesenchymal transition, Signal pathway

中图分类号:

R737.33

申晓畅,孙一卿,颜磊,赵兴波. 芳基烃受体核转位因子样蛋白2在子宫内膜癌中的表达[J]. 山东大学学报 (医学版), 2022, 60(5): 74-80.

SHEN Xiaochang, SUN Yiqing, YAN Lei, ZHAO Xingbo. Expression of aryl hydrocarbon receptor nuclear translocator-like 2 in endometrial cancer[J]. Journal of Shandong University (Health Sciences), 2022, 60(5): 74-80.

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