Zhang Yongya, Wu Fei, Guo Sibei, Yin Ruijie, Yuan Min, Li Xue, Zhao Xueru, Li Xiaohua
People's Hospital of Zhengzhou University, Zhengzhou, China.
Henan Key Laboratory of Ophthalmology and Visual Science, Henan Eye Institute, Henan Provincial People's Hospital, Henan Eye Hospital, Zhengzhou, China.
J Cell Physiol. 2024 Dec;239(12):e31429. doi: 10.1002/jcp.31429. Epub 2024 Sep 5.
Proliferative vitreoretinopathy (PVR) is a complex disease that significantly contributes to recurrent retinal detachment. Its development is notably affected by epithelial-mesenchymal transition (EMT), where apoptosis plays a crucial role as a regulator of EMT. However, the function of MeCP2 in governing apoptosis and EMT in retinal pigment epithelial (RPE) cells and its implications for PVR development have remained inadequately understood. Thus, we investigated the impact of MeCP2 on proliferation, migration, apoptosis and EMT in ARPE-19 cells to provide a fresh perspective on the etiology of PVR. The morphological changes in ARPE-19 cells induced by recombinant human MeCP2 protein and MeCP2 knockdown were observed. Wound healing assay were performed to verify the effects of recombinant human MeCP2 protein and MeCP2 knockdown on ARPE-19 cell migration. Furthermore, cell proliferation was assessed using the CCK-8 assay and flow cytometry. Western blot analysis, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), and immunofluorescence analysis were conducted to measure the protein levels associated with apoptosis, cell cycle and EMT. Western blot analysis and immunofluorescence assays confirmed that MeCP2 promoted EMT formation in ARPE-19 cells. The CCK-8 assay revealed that MeCP2 treatment enhanced the proliferation of ARPE-19 cells, whereas MeCP2 knockdown inhibited ARPE-19 cell proliferation. Treatment with recombinant human MeCP2 protein and MeCP2 knockdown altered the morphology of ARPE-19 cells. Wound healing assay demonstrated that MeCP2 knockdown inhibited ARPE-19 cell migration, and MeCP2 treatment promoted ARPE-19 cell migration. MeCP2 knockdown induced a G0/G1 phase block, inhibiting cell growth, and qRT-PCR data indicated reduced expression of cell cycle-related genes. Increased apoptosis was observed after MeCP2 knockdown in ARPE-19 cells. Overall, MeCP2 treatment stimulates cell proliferation, migration and EMT formation; conversely, MeCP2 knockdown inhibits EMT, cell proliferation, migration and cell cycle G1/S phase transition, and induces apoptosis.
增殖性玻璃体视网膜病变（PVR）是一种复杂疾病，对视网膜反复脱离有显著影响。其发展明显受上皮-间质转化（EMT）影响，其中细胞凋亡作为EMT的调节因子发挥关键作用。然而，甲基化CpG结合蛋白2（MeCP2）在视网膜色素上皮（RPE）细胞中调控细胞凋亡和EMT的功能及其对PVR发展的影响仍未得到充分了解。因此，我们研究了MeCP2对ARPE-19细胞增殖、迁移、凋亡和EMT的影响，为PVR的病因提供新视角。观察了重组人MeCP2蛋白和MeCP2基因敲低诱导的ARPE-19细胞形态变化。进行划痕愈合试验以验证重组人MeCP2蛋白和MeCP2基因敲低对ARPE-19细胞迁移的影响。此外，使用CCK-8试验和流式细胞术评估细胞增殖。进行蛋白质印迹分析、定量逆转录聚合酶链反应（qRT-PCR）和免疫荧光分析以测量与细胞凋亡、细胞周期和EMT相关的蛋白质水平。蛋白质印迹分析和免疫荧光试验证实MeCP2促进ARPE-19细胞中EMT的形成。CCK-8试验显示，MeCP2处理增强了ARPE-19细胞的增殖，而MeCP2基因敲低则抑制了ARPE-19细胞的增殖。重组人MeCP2蛋白处理和MeCP2基因敲低改变了ARPE-19细胞的形态。划痕愈合试验表明，MeCP2基因敲低抑制了ARPE-19细胞的迁移，而MeCP2处理促进了ARPE-19细胞的迁移。MeCP2基因敲低诱导G0/G1期阻滞，抑制细胞生长，qRT-PCR数据表明细胞周期相关基因的表达降低。在ARPE-19细胞中MeCP2基因敲低后观察到细胞凋亡增加。总体而言，MeCP2处理刺激细胞增殖、迁移和EMT形成；相反，MeCP2基因敲低抑制EMT、细胞增殖、迁移和细胞周期G1/S期转换，并诱导细胞凋亡。
