Li Xiaolong, Huang Yingwen, Guo Shunli, Xie Meiyu, Bin Xiaoyun, Shi Mingxia, Chen Anning, Chen Siyu, Wu Fan, Hu Qiping, Zhou Sufang
Department of Cell Biology and Genetics, School of Pre-Clinical Medicine, Guangxi Medical University, Nanning, Guangxi 530000, P.R. China.
Department of Biochemistry and Molecular Biology, School of Pre-Clinical Medicine, Guangxi Medical University, Nanning, Guangxi 530000, P.R. China.
Oncol Lett. 2019 Jul;18(1):609-616. doi: 10.3892/ol.2019.10374. Epub 2019 May 20.
Cervical adenocarcinoma (CA) is a type of cervical cancer, and in previous decades its incidence has steadily increased. The upregulation of regucalcin (RGN) in various tumor cell types inhibits the progression of cancer. To understand the role of RGN in CA, RGN expression in human cervical cancer compared with normal tissues was analyzed using The Cancer Genome Atlas database (TCGA). Subsequently, transfection of lentivirus-mediated RGN into HeLa cells was conducted to study its function in tumor proliferation and metastasis. The expression of RGN and proteins associated with the Wnt/β-catenin signaling pathway and epithelial-mesenchymal transition (EMT) were determined using reverse transcription-quantitative polymerase chain reaction and western blotting. Cell migration and invasion were evaluated using Transwell assays. Furthermore, cell proliferation, colony formation and cell cycle were assessed using the Cell Counting Kit-8, colony formation assay and flow cytometry, respectively. Lentivirus-mediated RGN effectively upregulated RGN expression, inhibited cell proliferation, retarded cellular invasion and promoted cell cycle arrest at the G/M phase in HeLa cells. In addition, the expression levels of β-catenin, p-glycogen synthase kinase (GSK)-3β, matrix metalloproteinase (MMP)-3, MMP-7 and MMP-9 were effectively decreased, whilst those of E-cadherin and GSK-3β were increased. The results suggest that RGN may be an inhibitory factor in tumorigenesis, and its mechanism of inhibiting tumor proliferation and metastasis may be associated with Wnt/β-catenin signaling and EMT.
宫颈腺癌（CA）是宫颈癌的一种类型，在过去几十年中其发病率稳步上升。在各种肿瘤细胞类型中，钙网蛋白（RGN）的上调可抑制癌症进展。为了解RGN在CA中的作用，利用癌症基因组图谱数据库（TCGA）分析了人宫颈癌与正常组织中RGN的表达。随后，将慢病毒介导的RGN转染到HeLa细胞中，以研究其在肿瘤增殖和转移中的功能。使用逆转录定量聚合酶链反应和蛋白质印迹法测定RGN以及与Wnt/β-连环蛋白信号通路和上皮-间质转化（EMT）相关的蛋白质的表达。使用Transwell实验评估细胞迁移和侵袭能力。此外，分别使用细胞计数试剂盒-8、集落形成实验和流式细胞术评估细胞增殖、集落形成和细胞周期。慢病毒介导的RGN有效上调了HeLa细胞中RGN的表达，抑制细胞增殖，阻碍细胞侵袭，并促进细胞周期停滞在G/M期。此外，β-连环蛋白、磷酸化糖原合酶激酶（GSK）-3β、基质金属蛋白酶（MMP）-3、MMP-7和MMP-9的表达水平有效降低，而E-钙黏蛋白和GSK-3β的表达水平升高。结果表明，RGN可能是肿瘤发生中的一种抑制因子，其抑制肿瘤增殖和转移的机制可能与Wnt/β-连环蛋白信号通路和EMT有关。
