Regulation of expression of HGF in BM‐MSCs by baculovirus‐mediated transduction
Regulation of expression of HGF in BM‐MSCs by baculovirus‐mediated transduction
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DOI：
10.1002/cbin.10071
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发表时间：
2013-07
影响因子：
3.9
通讯作者：
Yi Ji Tu;Aifang Ye;Zhi-min Pan;Chao Zheng;Tian-Long Wu;Xi-gao Cheng;Fei Guo
中科院分区：
文献类型：
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作者：
Yi Ji Tu;Aifang Ye;Zhi-min Pan;Chao Zheng;Tian-Long Wu;Xi-gao Cheng;Fei Guo
近年来，骨髓间充质干细胞（BM-MSCs）移植被广泛用于治疗股骨头坏死（ONFH）。此外，众所周知，由于HGF强大的血管生成和抗纤维化能力，将肝细胞生长因子（HGF）引入BM-MSC中将大大提高干细胞治疗的治疗效果。然而，HGF在体内持续过度表达可能会导致肉瘤，例如卡波西肉瘤。为了增强HGF修饰干细胞移植对ONFH的治疗效果并防止其副作用，我们试图构建一个基因调控系统来严格、准确地控制BM-MSC中HGF的表达。我们选择杆状病毒作为基因载体，并引入pTet-on先进系统。最后，成功构建了病毒载体vAcrtTA2s-Ptight-HGF，并将其转入BM-MSCs以调控HGF的准确表达。结果显示，不同诱导剂量的强力霉素（DOX）通过ELISA和Western blot验证了不同水平的HGF表达。它们之间存在剂量反应关系，DOX体外诱导BM-MSCs HGF表达的最佳剂量为1 µg/mL。我们的结论是，通过杆状病毒介导的一次性转导来调节 BM-MSC 中 HGF 的表达是可行的。
Bone marrow‐derived mesenchymal stem cells (BM‐MSCs) transplantation is widely adopted for the curing of osteonecrosis of femoral head (ONFH) in recent years. Furthermore, it is known that introducing hepatocyte growth factor (HGF) into BM‐MSCs will greatly improve the therapeutic effect of stem‐cell therapy owing to the great angiogenic and anti‐fibrotic capabilities of HGF. However, continuing overexpression of HGF in vivo may cause sarcomas, such as Kaposi's sarcoma. Aiming at enhancing the therapeutic effect and preventing the side effects of HGF‐modified stem‐cell transplantation on ONFH, we sought to construct a gene regulation system to control HGF expression in BM‐MSCs rigorously and accurately. We selected baculovirus as the gene vector and introduced pTet‐on advanced system into that. Finally, a virus vector vAcrtTA2s‐Ptight‐HGF was successfully built and delivered into BM‐MSCs to regulate the accurate expression of HGF. As shown in the results, different levels of HGF expression were verified by ELISA and Western blot with different induction doses of doxycycline (DOX). There was a dose–response relationship between them, and the optimum dose of DOX to induce HGF expression in BM‐MSCs in vitro was 1 µg/mL. We conclude that it is feasible to regulate HGF expression in BM‐MSCs by baculovirus‐mediated one‐off transduction.