Regulatory dynamics of Nanog in chondrocyte dedifferentiation: role of KLF4/p53 and p38/AKT signaling

IF 3.9 4区 生物学 Q1 GENETICS & HEREDITY
Young Seok Eom, Song Ja Kim
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Abstract

Homeobox protein Nanog, a member of the transcription factor family, plays a crucial role in maintaining the pluripotency and self-renewal of embryonic stem cells. Due to its diverse activities, Nanog has been identified in multiple cell types, including embryonic stem cells (ESCs) and cancer stem cells (CSCs). However, its molecular mechanism in chondrocytes remains unclear. In this study, we explored the effects of Nanog on chondrocytes and its interaction with chondrocyte-specific proteins. Chondrocytes were transfected with a Nanog cDNA vector, resulting in reduced expression of the chondrogenic markers Type II collagen and SOX9, as confirmed by western blot, RT-PCR, and immunofluorescence. Following siRNA transfection, the dedifferentiation effect of Nanog was reversed, restoring Type II collagen and SOX9 expression. We also discovered that the mechanism by which Nanog affects chondrocytes is closely linked to p53 and KLF4. Overexpression of KLF4 induced the phosphorylation of p53, and phospho-p53 directly inhibited Nanog expression. Moreover, the p53 activator Nutlin-3 A accelerated Nanog degradation, while the p53 inhibitor Pifithrin-α stabilized Nanog. Stabilized Nanog continued to promote chondrocyte dedifferentiation. Additional experiments were performed to identify the signaling pathways involved in Nanog-induced chondrocyte dedifferentiation. Our results showed that Nanog overexpression in chondrocytes significantly impacted the p38 kinase and AKT signaling pathways. Inhibition of p38 and AKT with SB203580 and LY294002 reduced Nanog expression and partially restored Type II collagen levels. Conversely, activation with anisomycin(ANS) and 740 Y-P enhanced Nanog expression, further reducing Type II collagen levels. To investigate Nanog’s role in early development in vivo, we injected Nanog expression vectors into zebrafish embryos. The injected zebrafish exhibited structural defects in craniofacial cartilage, confirming Nanog’s involvement in chondrocyte differentiation. These findings suggest that Nanog induces chondrocyte dedifferentiation, and this process can be modulated via the p53/KLF4 and p38/AKT pathways.

Abstract Image

Nanog在软骨细胞去分化中的调控动力学:KLF4/p53和p38/AKT信号的作用
同源盒蛋白Nanog是转录因子家族的一员,在维持胚胎干细胞的多能性和自我更新中起着至关重要的作用。由于其多种活性,Nanog已在多种细胞类型中被发现,包括胚胎干细胞(ESCs)和癌症干细胞(CSCs)。然而,其在软骨细胞中的分子机制尚不清楚。在这项研究中,我们探讨了Nanog对软骨细胞的影响及其与软骨细胞特异性蛋白的相互作用。用Nanog cDNA载体转染软骨细胞后,经western blot、RT-PCR和免疫荧光检测证实,软骨形成标志物II型胶原和SOX9的表达减少。转染siRNA后,Nanog的去分化作用被逆转,恢复II型胶原和SOX9的表达。我们还发现Nanog影响软骨细胞的机制与p53和KLF4密切相关。KLF4过表达诱导p53磷酸化,磷酸化p53直接抑制Nanog的表达。此外,p53激活剂Nutlin-3 A加速了Nanog的降解,而p53抑制剂Pifithrin-α则稳定了Nanog。稳定的Nanog继续促进软骨细胞去分化。我们还进行了其他实验,以确定参与纳米颗粒诱导的软骨细胞去分化的信号通路。我们的研究结果表明,Nanog在软骨细胞中的过表达显著影响p38激酶和AKT信号通路。SB203580和LY294002抑制p38和AKT可降低Nanog表达,部分恢复II型胶原水平。相反,用anisomycin(ANS)和740 Y-P激活可以增强Nanog的表达,进一步降低II型胶原蛋白的水平。为了研究Nanog在体内早期发育中的作用,我们将Nanog表达载体注入斑马鱼胚胎。注射后的斑马鱼颅面软骨出现结构性缺陷,证实Nanog参与软骨细胞分化。这些发现表明Nanog诱导软骨细胞去分化,这一过程可以通过p53/KLF4和p38/AKT通路调节。
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来源期刊
CiteScore
3.50
自引率
3.40%
发文量
92
审稿时长
2 months
期刊介绍: Functional & Integrative Genomics is devoted to large-scale studies of genomes and their functions, including systems analyses of biological processes. The journal will provide the research community an integrated platform where researchers can share, review and discuss their findings on important biological questions that will ultimately enable us to answer the fundamental question: How do genomes work?
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