增生性玻璃体视网膜病变(PVR)是一种复杂的疾病,显着有助于复发性视网膜脱离。它的发育受到上皮-间质转化(EMT)的显著影响,其中凋亡作为EMT的调节剂起着至关重要的作用。然而,MeCP2在调节视网膜色素上皮(RPE)细胞凋亡和EMT中的功能及其对PVR发展的影响仍未得到充分了解。因此,我们研究了MeCP2对增殖的影响,迁移,ARPE-19细胞的凋亡和EMT为PVR的病因提供了新的视角。观察重组人MeCP2蛋白和MeCP2敲低诱导ARPE-19细胞的形态学变化。进行伤口愈合测定以验证重组人MeCP2蛋白和MeCP2敲低对ARPE-19细胞迁移的影响。此外,使用CCK-8测定和流式细胞术评估细胞增殖。蛋白质印迹分析,定量逆转录酶-聚合酶链反应(qRT-PCR),和免疫荧光分析进行测量与细胞凋亡相关的蛋白质水平,细胞周期和EMT。Western印迹分析和免疫荧光测定证实MeCP2促进ARPE-19细胞中的EMT形成。CCK-8检测显示MeCP2处理增强了ARPE-19细胞的增殖,而MeCP2敲低抑制ARPE-19细胞增殖。用重组人MeCP2蛋白和MeCP2敲低处理改变了ARPE-19细胞的形态。伤口愈合实验证明MeCP2敲低抑制ARPE-19细胞迁移,和MeCP2处理促进ARPE-19细胞迁移。MeCP2敲低诱导G0/G1期阻滞,抑制细胞生长,和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敲低后观察到细胞凋亡增加。总的来说,MeCP2处理刺激细胞增殖,迁移和EMT形成;相反,MeCP2敲低抑制EMT,细胞增殖,迁移和细胞周期G1/S相变,并诱导细胞凋亡。
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.