Dystrophin Dp71ab is monoclonally expressed in human satellite cells and enhances proliferation of myoblast cells.
Dystrophin Dp71ab is monoclonally expressed in human satellite cells and enhances proliferation of myoblast cells.
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Dystrophin Dp71ab在人卫星细胞中单克隆表达并增强成肌细胞的增殖。
DOI：
10.1038/s41598-020-74157-y
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发表时间：
2020-10-13
影响因子：
4.6
通讯作者：
Matsuo M
中科院分区：
文献类型：
--
作者：
Farea M;Rani AQM;Maeta K;Nishio H;Matsuo M
肌营养不良蛋白 Dp71 是 DMD 基因的最小亚型，其突变会导致杜氏肌营养不良症 (DMD)。 Dp71 也已被证明在各种细胞过程中发挥作用。基于干细胞的疗法可能有效治疗 DMD，但无法产生足够数量的干细胞仍然是一个重大障碍。尽管 Dp71 由许多变体组成，但卫星细胞中的 Dp71 尚未得到研究。在此，通过逆转录-PCR从人类卫星细胞的总RNA中扩增出由外显子G1至79的18个外显子组成的全长Dp71。扩增产物显示所有测序克隆中外显子 71 和 78 均缺失，表明 Dp71ab 的单克隆表达。卫星细胞裂解物的蛋白质印迹显示与过度表达的 Dp71ab 相对应的条带。与转染模拟质粒的细胞相比，将表达 Dp71ab 的质粒转染至人成肌细胞中显着增强了细胞增殖。然而，编码所有 18 个外显子的 Dp71 表达质粒的转染并没有增强成肌细胞增殖。这些发现表明，Dp71ab（而非 Dp71）是成肌细胞增殖的分子增强剂，并且用 Dp71ab 转染可能会产生用于 DMD 治疗的高产量干细胞。
Dystrophin Dp71 is the smallest isoform of the DMD gene, mutations in which cause Duchenne muscular dystrophy (DMD). Dp71 has also been shown to have roles in various cellular processes. Stem cell-based therapy may be effective in treating DMD, but the inability to generate a sufficient number of stem cells remains a significant obstacle. Although Dp71 is comprised of many variants, Dp71 in satellite cells has not yet been studied. Here, the full-length Dp71 consisting of 18 exons from exons G1 to 79 was amplified by reverse transcription-PCR from total RNA of human satellite cells. The amplified product showed deletion of both exons 71 and 78 in all sequenced clones, indicating monoclonal expression of Dp71ab. Western blotting of the satellite cell lysate showed a band corresponding to over-expressed Dp71ab. Transfection of a plasmid expressing Dp71ab into human myoblasts significantly enhanced cell proliferation when compared to the cells transfected with the mock plasmid. However, transfection of the Dp71 expression plasmid encoding all 18 exons did not enhance myoblast proliferation. These findings indicated that Dp71ab, but not Dp71, is a molecular enhancer of myoblast proliferation and that transfection with Dp71ab may generate a high yield of stem cells for DMD treatment.
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影响因子：
2.5
作者：
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通讯作者：
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影响因子：
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作者：
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3.1
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通讯作者：
Dilworth, F. Jeffrey
DOI：
10.1016/j.bbrc.2018.11.168
发表时间：
2019-01-08
影响因子：
3.1
作者：
Rani, Abdul Qawee Mahyoob;Farea, Manal;Matsuo, Masafumi
通讯作者：
Matsuo, Masafumi