Wang Jing, He Lijie, Han Zhe
Department of Clinical Laboratory, Tianjin Fifth Central Hospital, Tianjin 300450, China.
Department of Clinical Laboratory, Tianjin Fifth Central Hospital, Tianjin 300450, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2024 Dec;40(12):1089-1095.
Objective The purpose of this study was to investigate how miR-200b-3p inhibitors the proliferation and metastasis of endometrial cancer(EC) cells by inducing the expression of FOS-like antigen 2(FOSL2) of activator protein 1(AP1) transcription family. Methods Endometrial cancer cell line HEC-1-A was divided into 12 groups: NC-mimic (transfected with negative control NC mimic), miR-200b-3p mimic (transfected with miR-200b-3p mimic), NC-inhibitor (transfected with negative control NC inhibitor), miR-200b-3p inhibitor group (transfected with miR-200b-3p inhibitor), si-NC (transfected with negative control Si-NC), si-FOSL2 (transfected with si-FOSL2), oe-NC (transfected with negative control oe-NC), oe-FOSL2 group (oe-FOSL2), miR-200b-3p mimic+oe-NC group (co-transfected with miR-200b-3p mimic and oe-NC), miR-200b-3p mimic+oe-FOSL2 group (co-transfected with miR-200b-3p mimic and oe-FOSL2), miR-200b-3p inhibitor+si-NC group (co-transfected with miR-200b-3p inhibitor and si-NC), miR-200b-3p inhibitor+si-FOSL2 group (co-transfected with miR-200b-3p inhibitor and si-FOSL2). Real-time fluorescence quantitative PCR, Western blot, CCK-8 assay, scratch test and Transwell assay were used to detect the expression of miR-200b-3p mRNA, FOSL2 mRNA and protein expression level, cell proliferation, migration and invasion. Results In endometrial cancer cell lines, the expression of miR-200b-3p was significantly down-regulated, while the expression of FOSL2 was significantly up-regulated. Compared with NC-mimic group, the expression of FOSL2, N-cadherin and Vimentin in miR-200b-3p mimic group was significantly decreased, and the expression of E-cadherin was significantly increased. The cell proliferation, migration rate and the number of transmembrane cells were significantly decreased. Compared with the miR-200b-3p mimic+oe-NC group, the expression of FOSL2, N-cadherin and Vimentin in miR-200b-3p mimic+oe-FOSL2 group was significantly increased, and the expression level of E-cadherin was significantly decreased, and the cell proliferation, migration rate and the number of transmembrane cells were significantly increased. Compared with NC-inhibitor group, the expression of FOSL2, N-cadherin and Vimentin in miR-200b-3p inhibitor group was significantly increased, and the expression of E-cadherin was significantly decreased. The cell proliferation, migration rate and the number of transmembrane cells were significantly increased. Compared with the miR-200b-3p inhibitor+si-NC group, the expression of FOSL2, N-cadherin and Vimentin in miR-200b-3p inhibitor+si-FOSL2 group was significantly decreased, and the expression of E-cadherin was significantly increased; the cell proliferation, migration rate and the number of transmembrane cells were significantly decreased. Conclusion The expression of miR-200b-3p in endometrial cancer cells is down-regulated, which can inhibitor the proliferation, migration and invasion of endometrial cancer cells by regulating the EMT process, and its mechanism is related to its targeted negative regulation of FOSL2 expression.
目的 本研究旨在探讨miR-200b-3p如何通过诱导激活蛋白1(AP1)转录家族的FOS样抗原2(FOSL2)表达来抑制子宫内膜癌(EC)细胞的增殖和转移。方法 将子宫内膜癌细胞系HEC-1-A分为12组:NC-mimic(转染阴性对照NC mimic)、miR-200b-3p mimic(转染miR-200b-3p mimic)、NC-inhibitor(转染阴性对照NC inhibitor)、miR-200b-3p inhibitor组(转染miR-200b-3p inhibitor)、si-NC(转染阴性对照Si-NC)、si-FOSL2(转染si-FOSL2)、oe-NC(转染阴性对照oe-NC)、oe-FOSL2组(oe-FOSL2)、miR-200b-3p mimic+oe-NC组(共转染miR-200b-3p mimic和oe-NC)、miR-200b-3p mimic+oe-FOSL2组(共转染miR-200b-3p mimic和oe-FOSL2)、miR-200b-3p inhibitor+si-NC组(共转染miR-200b-3p inhibitor和si-NC)、miR-200b-3p inhibitor+si-FOSL2组(共转染miR-200b-3p inhibitor和si-FOSL2)。采用实时荧光定量PCR、蛋白质免疫印迹法、CCK-8法、划痕试验和Transwell试验检测miR-200b-3p mRNA、FOSL2 mRNA表达及蛋白表达水平、细胞增殖、迁移和侵袭能力。结果 在子宫内膜癌细胞系中,miR-200b-3p表达显著下调,而FOSL2表达显著上调。与NC-mimic组相比,miR-200b-3p mimic组FOSL2、N-钙黏蛋白和波形蛋白表达显著降低,E-钙黏蛋白表达显著增加。细胞增殖、迁移率和穿膜细胞数显著降低。与miR-200b-3p mimic+oe-NC组相比,miR-200b-3p mimic+oe-FOSL2组FOSL2、N-钙黏蛋白和波形蛋白表达显著增加,E-钙黏蛋白表达水平显著降低,细胞增殖、迁移率和穿膜细胞数显著增加。与NC-inhibitor组相比,miR-200b-3p inhibitor组FOSL2、N-钙黏蛋白和波形蛋白表达显著增加,E-钙黏蛋白表达显著降低。细胞增殖、迁移率和穿膜细胞数显著增加。与miR-200b-3p inhibitor+si-NC组相比,miR-200b-3p inhibitor+si-FOSL2组FOSL2、N-钙黏蛋白和波形蛋白表达显著降低,E-钙黏蛋白表达显著增加;细胞增殖、迁移率和穿膜细胞数显著降低。结论 子宫内膜癌细胞中miR-200b-3p表达下调,其可通过调节EMT过程抑制子宫内膜癌细胞的增殖、迁移和侵袭,其机制与其对FOSL2表达的靶向负调控有关。