Forced expression of mouse progerin attenuates the osteoblast differentiation interrupting beta-catenin signal pathway in vitro.
Forced expression of mouse progerin attenuates the osteoblast differentiation interrupting beta-catenin signal pathway in vitro.
复制标题
小鼠早老蛋白的强制表达会减弱成骨细胞的分化，从而在体外中断β-连环蛋白信号通路。
DOI：
10.1007/s00441-018-2930-y
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发表时间：
2019
期刊：
影响因子：
3.6
通讯作者：
Takahashi Tomihisa
中科院分区：
文献类型：
--
作者：
Tsukune Naoya;Naito Masako;Ohashi Akiko;Ninomiya Tadashi;Sato Shuichi;Takahashi Tomihisa
核蛋白laminA是核质内膜的组成部分，在核形成和细胞分化中起重要作用。突变的核纤层蛋白A（称为早老蛋白）的表达导致罕见的遗传性衰老疾病，即Hutchinson-Gilford早老综合征，其表现为骨形成异常，成骨细胞和骨细胞数量减少。然而，早老素抑制成骨的确切分子机制尚未完全阐明。在这里，我们创建了小鼠核纤层蛋白A dC 50 cDNA编码的早老蛋白，缺乏50个氨基酸残基的C-末端，转染在小鼠前成骨细胞样MC 3 T3-E1细胞，并检查成骨细胞表型的变化。当核纤层蛋白A dC 50表达的细胞与分化诱导培养基一起培养时，碱性磷酸酶（ALP）活性和主要成骨细胞标志物I型胶原（Col 1）、骨唾液蛋白（BSP）、牙本质基质蛋白1（DMP 1）和Runx 2的mRNA水平显著降低，并且没有检测到矿化结节，如在表达空载体的对照细胞中所见。在矿化诱导培养基的培养中，BSP，骨钙素，DMP 1，Runx 2和osterix的mRNA水平强烈下降，与核纤层蛋白A dC 50表达细胞的矿化损失平行，而矿化结节出现在21天的对照细胞。此外，随着GSK-3β磷酸化水平的降低，核纤层蛋白A dC 50表达降低，β-catenin核定位降低。这些结果表明，核纤层蛋白A dC 50抑制成骨细胞分化的早期和晚期阶段，它负性调节β-catenin活性与GSK-3β在细胞质中的相互作用。
Nuclear protein, lamin A, which is a component of inner membrane on nucleoplasm, plays a role in nuclear formation and cell differentiation. The expression of mutated lamin A, termed progerin, causes a rare genetic aging disorder, Hutchinson-Gilford progeria syndrome, which shows abnormal bone formation with the decrease in a number of osteoblasts and osteocytes. However, exact molecular mechanism how progerin exerts depressive effects on osteogenesis has not been fully understood. Here, we created mouse lamin A dC50 cDNA encoding progerin that lacks 50 amino acid residues at C-terminus, transfected it in mouse preosteoblast-like MC3T3-E1 cells, and examined the changes in osteoblast phenotype. When lamin A dC50-expressed cells were cultured with differentiation-inductive medium, alkaline phosphatase (ALP) activity and mRNA levels of major osteoblast markers, type I collagen (Col1), bone sialoprotein (BSP), dentine matrix protein 1 (DMP1), and Runx2 were significantly decreased, and no mineralized nodules were detected as seen in control cells expressing empty vector. In the culture with mineralization-inductive medium, mRNA levels of BSP, osteocalcin, DMP1, Runx2, and osterix were strongly decreased parallel with loss of mineralization in lamin A dC50-expressed cells, while mineralized nodules appear at 21 days in control cells. Furthermore, lamin A dC50 expression was depressed nuclear localization of β-catenin with the decrease of GSK-3β phosphorylation level. These results suggest that lamin A dC50 depresses osteoblast differentiation in both early and late stages, and it negatively regulates β-catenin activity interacting with GSK-3β in cytoplasm.