目的：探讨特异性激活G蛋白偶联雌激素受体（GPER）对结直肠癌（CRC）细胞迁移的作用及其可能的机制。方法：体外培养CRC细胞RKO和SW480，使用0.5或1.0 μmol/L 的GPER特异性激活剂（G-1）处理CRC细胞，采用CCK-8法、划痕实验和Transwell 实验分别检测G-1对CRC细胞增殖、迁移的影响。用qPCR和WB法分别检测G-1及G-1+活性氧（ROS）清除剂N-乙酰半胱氨酸（NAC）处理对RKO细胞E-钙黏素（E-Cad）、纤连蛋白（FN）mRNA和蛋白表达的影响，用流式细胞术检测RKO细胞中ROS的水平。结果：经G-1处理后RKO和SW480 细胞的迁移能力均明显减弱（均P<0.05），能显著上调细胞中E-cad mRNA及蛋白表达、下调FN mRNA及蛋白表达（均P<0.05）。G-1处理能刺激RKO 细胞ROS 水平上升，在NAC的作用下，由G-1引起的E-cad、FN蛋白表达变化被部分逆转。结论：特异性激活GPER通过上调ROS水平抑制EMT进程，进而抑制CRC细胞的迁移。

Objective: To investigate the impact of specific activation of G protein-coupled estrogen receptors (GPER) on the migration of colorectal cancer (CRC) cells, and explore the potential underlying mechanism. Methods: CRC RKO and SW480 cells were cultured in vitro and treated with 0.5 or 1.0 μmol/L GPER-specific activator (G-1). CCK-8 method, scratch test, and Transwell test were used to detect the effects of G-1 on CRC cell proliferation and migration. The impact of G-1/G-1+ reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC) treatment on the mRNA and protein expressions of E-cadherin (E-Cad) and fibronectin (FN) in RKO cells was measured using qPCR and WB. The level of ROS in RKO cells was detected by flow cytometry. Results: The migration ability of RKO and SW480 cells after the treatment with G-1 was significantly reduced (all P<0.05). Additionally, G-1 treatment significantly increased the mRNA and protein expressions of E-cad and decreased the mRNA and protein expressions of FN in the cells (all P<0.05). G-1 treatment also stimulated the production of ROS in RKO cells. However, the protein expression changes of E-cad and FN caused by G-1 were partially reversed under the action of NAC. Conclusion: Speicific activation of GPER inhibits the migration of CRC cells by suppressing epithelial-mesenchymal transition (EMT). This inhibitory effect is likely mediated through the upregulation of ROS levels in CRC cells.

重庆市自然科学基金项目（No. cstc2020jcyj-msxmX0117）；重庆市教委科学技术研究项目（No. KJQN202102802）；重庆医药高等专科学校自然科学技术研究项目（No. ygz2018107, ygz2021123）