Suppression of lamin A/C by short hairpin RNAs promotes adipocyte lineage commitment in mesenchymal progenitor cell line, ROB-C26
Suppression of lamin A/C by short hairpin RNAs promotes adipocyte lineage commitment in mesenchymal progenitor cell line, ROB-C26
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DOI：
10.1007/s00418-011-0890-3
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发表时间：
2012-02-01
影响因子：
2.3
通讯作者：
Takahashi, Tomihisa
中科院分区：
文献类型：
--
作者：
Naito, Masako;Omoteyama, Kazuki;Takahashi, Tomihisa
核纤层蛋白A/C基因编码一种核膜蛋白，该基因的突变与多种退行性疾病有关，这些疾病与过早衰老有关。虽然核纤层蛋白A/C参与组织稳态的调节，但在分化为脂肪细胞的间充质细胞中的不同表达模式知之甚少。在这里，我们研究了核纤层蛋白A/C在大鼠间充质祖细胞系ROB-C26（C26）中的表达。免疫细胞化学分析显示，核纤层蛋白A/C在未成熟脂肪细胞中表达短暂下调，但其表达随着终末分化而增加。为了阐明核纤层蛋白A/C的表达对间充质细胞分化的作用，核纤层蛋白A/C的表达被抑制使用短发夹RNA（shRNA）分子在C26细胞。在缺乏脂肪形成刺激的情况下，核纤层蛋白A/C shRNA降低碱性磷酸酶（ALP）活性，但诱导前脂肪细胞因子-1（Pref-1）mRNA表达。在脂肪形成刺激物的存在下，核纤层蛋白A/C敲低促进脂肪细胞分化，如通过检测油红O染色的增加所评估的。RT-PCR分析显示，核纤层蛋白A/C shRNA可诱导脂肪细胞分化过程中PPAR γ 2和aP 2 mRNA表达增加。这些结果表明，降低核纤层蛋白A/C的表达水平，不仅抑制成骨细胞表型，但也促进脂肪细胞分化的C26细胞。
Lamin A/C gene encodes a nuclear membrane protein, and mutations in this gene are associated with diverse degenerative diseases that are linked to premature aging. While lamin A/C is involved in the regulation of tissue homeostasis, the distinct expression patterns are poorly understood in the mesenchymal cells differentiating into adipocytes. Here, we examined the expression of lamin A/C in a rat mesenchymal progenitor cell-line, ROB-C26 (C26). Immunocytochemical analysis showed that lamin A/C was transiently down-regulated in immature adipocytes, but its expression increased with terminal differentiation. To elucidate the role of lamin A/C expression on mesenchymal cell differentiation, lamin A/C expression was suppressed using short hairpin RNA (shRNA) molecules in C26 cells. In the absence of adipogenic stimuli, lamin A/C shRNA decreased alkaline phosphatase (ALP) activity, but induced preadipocyte factor -1 (Pref-1) mRNA expression. In the presence of adipogenic stimuli, lamin A/C knockdown promotes adipocytes differentiation, as assessed by the detection of an increase in Oil Red O staining. RT-PCR analysis showed that lamin A/C shRNA resulted in increased mRNA expression of PPAR gamma 2 and aP2 during adipocyte differentiation. These results suggest that decreased lamin A/C expression levels not only suppress osteoblast phenotypes but also promote adipocyte differentiation in C26 cells.