MIF缺失抑制子宫内膜癌细胞的增殖转移及机制研究

Study on the inhibitory effect and mechanism of MIF deficiency on the proliferation and metastasis of endometrial cancer cells

摘要

目的 探讨巨噬细胞迁移抑制因子(MIF)缺失对子宫内膜癌(EC)细胞的转移以及细胞铁死亡的影响及调节机制.方法 利用蛋白质免疫印迹检测人子宫内膜内皮细胞(hEEC)与EC细胞系(Ishikawa、KLE、RL95-2与AN3-CA)中MIF的表达水平.将Ishikawa与KLE细胞各分为4组,即si-NC转染组(si-NC)、si-MIF转染组(si-MIF)、si-MIF+oe-NC转染组(si-MIF+oe-NC)与si-MIF+oe-SLC7A1转染组(si-MIF+oe-SLC7A1).蛋白质免疫印迹法测定各组细胞中MIF、SLC7A1、p-AKT/AKT与p-GSK-3β/GSK-3β水平;CCK-8法检测细胞活力;划痕试验检测细胞迁移;Transwell试验检测细胞侵袭;测定丙二醛(MDA)与Fe²⁺水平.结果 与hEEC组相比,Ishikawa、KLE、RL95-2与AN3-CA组细胞中MIF的表达量显著升高(F=23.933,P<0.001).与si-NC组相比,si-MIF与si-MIF+oe-NC组SLC7A1的表达量均显著降低(F=20.157,P<0.001),si-MIF与si-MIF+oe-NC组p-AKT/AKT与p-GSK-3β/GSK-3β水平、细胞增殖、迁移与侵袭能力均显著降低(F=12.456、18.234、32.451、12.257、15.364,P<0.001),MDA与Fe²⁺的水平显著升高(F=40.215、38.845,P<0.001).与si-MIF+oe-NC组相比,si-MIF+oe-SLC7A1组p-AKT/AKT与p-GSK-3β/GSK-3β水平、细胞增殖、迁移与侵袭能力均显著升高(F=12.397、21.045、26.229、18.330、14.228,P<0.001),MDA与Fe²⁺的水平显著降低(F=25.294、26.005,P<0.001).结论 MIF缺失可能通过抑制SLC7A1进而抑制AKT/GSK-3β信号通路,促进细胞铁死亡,抑制EC细胞增殖、迁移与侵袭.

Abstract

Objective To investigate the effects and regulatory mechanisms of macrophage migration inhibition factor (MIF) deficiency on metastasis and ferroptosis in endometrial cancer (EC) cells. Methods Western blot analysis was used to detect MIF expression levels in human endometrial endothelial cells (hEEC) and EC cell lines (Ishikawa, KLE, RL95-2 and AN3-CA). Ishikawa and KLE cells were divided into four groups: si-NC transfection group (si-NC), si-MIF transfection group (si-MIF), si-MIF+oe-NC transfection group (si-MIF+oe-NC), and si-MIF+oe-SLC7A1 transfection group (si-MIF+oe-SLC7A1). Western blotting was used to measure the levels of MIF, SLC7A1, p-AKT/AKT, and p-GSK-3β/GSK-3β in each group. Cell viability was assessed using the CCK-8 assay, while wound healing and Transwell assays were performed to evaluate cell migration and invasion, respectively. Additionally, malondialdehyde (MDA) and Fe ²⁺ levels were measured. Results Compared to the hEEC group, MIF expression was significantly upregulated in Ishikawa, KLE, RL95-2, and AN3-CA cells (F=23.933, P<0.001). Compared to the si-NC group, the si-MIF and si-MIF+oe-NC groups exhibited significantly reduced SLC7A1 expression (F=20.157, P<0.001). Furthermore, p-AKT/AKT and p-GSK-3β/GSK-3β levels, cell proliferation, migration, and invasion abilities were significantly decreased in the si-MIF and si-MIF+oe-NC groups (F=12.456, 18.234, 32.451, 12.257 and 15.364, respectively, P<0.001), while MDA and Fe ²⁺ levels were significantly increased (F=40.215 and 38.845, respectively, P<0.001). Compared to the si-MIF+oe-NC group, the si-MIF+oe-SLC7A1 group showed significantly increased p-AKT/AKT and p-GSK-3β/GSK-3β levels, as well as enhanced cell proliferation, migration, and invasion abilities (F=12.397, 21.045, 26.229, 18.330 and 14.228, respectively, P<0.001), while MDA and Fe ²⁺ levels were significantly decreased (F=25.294 and 26.005, respectively, P<0.001). Conclusion MIF deficiency may inhibit the AKT/GSK-3β signaling pathway by suppressing SLC7A1 expression, thereby promoting ferroptosis and inhibiting EC cell proliferation, migration, and invasion.

关键词

子宫内膜癌 / 巨噬细胞迁移抑制因子 / 信号通路 / 细胞转移 / 铁死亡

Key words

endometrial cancer / macrophage migration inhibition factor / signaling pathway / cell migration / ferroptosis

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基金资助

邯郸市科学技术研究与发展计划(19422083029ZC)