摘要
目的:探讨过表达KLF4通过Wnt/β-catenin信号通路抑制胃癌细胞上皮-间质转化和细胞周期的作用机制。方法:将课题组前期已成功转染过表达KLF4的BGC-823细胞分为KLF4过表达载体组(BGC823-OE组)、空病毒载体组(BGC823-NC组)和未经处理的BGC-823细胞组(BGC823-Ctrl组)。Western blot和qRT-PCR检测KLF4、EMT和Wnt信号通路相关蛋白和mRNA在三组细胞中的表达水平。CCK-8实验、划痕实验、Transwell迁移实验、平板克隆实验和流式细胞术观察各组细胞的增殖能力、迁移能力、克隆形成能力和细胞周期变化情况。结果:Western blot和qRT-PCR实验结果显示BGC823-OE组KLF4蛋白和mRNA表达水平明显升高。上皮-间质转化的上皮标志物E-cadherin蛋白和mRNA表达水平升高,间质标志物N-cadherin蛋白和mRNA表达水平降低。Wnt信号通路下游靶基因β-catenin和Cyclin D1的蛋白和mRNA表达水平降低。CCK-8增殖实验结果显示BGC823-OE组的增殖能力下降。划痕实验和Transwell迁移实验结果显示BGC823-OE组的迁移能力下降。平板克隆结果显示BGC823-OE组的克隆能力下降。流式细胞术结果显示BGC823-OE组G_(0)/G_(1)期比例增高,S期比例降低。结论:过表达KLF4可能通过Wnt/β-catenin信号通路抑制胃癌细胞EMT及阻滞胃癌细胞的细胞周期。
Objective:To investigate the mechanism by which overexpression of KLF4 inhibits epithelial-mesenchymal transition(EMT) and cell cycle in gastric cancer cells through the Wnt/β-catenin signaling pathway.Methods:BGC-823 cells that had been successfully transfected with KLF4 expression in the previous phase of the group were divided into a KLF4 overexpression vector group(BGC823-OE group),an empty virus vector group(BGC823-NC group) and an untreated BGC-823 cell group(BGC823-Ctrl group).Western blot and qRT-PCR were performed to detect the expression levels of KLF4,EMT and Wnt signalling pathway related proteins and mRNA in the three groups of cells.CCK-8 assay, scratch assay, Transwell migration assay, plate cloning assay and flow cytometry were used to observe the proliferation ability, migration ability, clone formation ability and cell cycle changes of each group of cells.Results:The results of Western blot and qRT-PCR experiments showed that the expression levels of KLF4 protein and mRNA were significantly increased in the BGC823-OE group.The epithelial marker E-cadherin protein and mRNA expression levels were increased and the mesenchymal marker, N-cadherin protein and mRNA expression levels were decreased for epithelial-mesenchymal transition.The protein and mRNA expression levels of β-catenin and Cyclin D1,the downstream targets of the Wnt signaling pathway were reduced.The results of the CCK-8 assay showed a reduced proliferative capacity in the BGC823-OE group.The results of the scratch assay and Transwell migration assay showed a decrease in the migration ability of the BGC823-OE group. The results of theplate cloning showed reduced cloning ability in the BGC823-OE group. Flow cytometry results showed an increasedproportion of G_(0)/G_(1)phase and a decreased proportion of S phase in the BGC823-OE group.Conclusion:Overex-pression of KLF4 may inhibit EMT and block the cell cycle of gastric cancer cells through the Wnt/β-catenin signa-ling pathway.
作者
符悦
李春鸣
梁娜
罗庆庆
吴丝
周航皓
FU Yue;LI Chunming;LIANG Na;LUO Qingqing;WU Si;ZHOU Hanghao(Zunyi Medical University,Guizhou Zunyi 563000,China;Department of Tissue and Embryology,Zunyi Medical University,Guizhou Zunyi 563000,China;Department of Pathology,Affiliated Hospital of Zunyi Medical University,Guizhou Zunyi 563000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第6期991-996,共6页
Journal of Modern Oncology
基金
贵州省研究生科研基金(编号:黔教合YJSKYJJ[2021]181)
贵州省遵义市科技局联合基金资助项目[编号:遵市科合HZ字(2020)52号]。
