MiR-199a-modified exosomes from adipose tissue-derived mesenchymal stem cells improve hepatocellular carcinoma chemosensitivity through mTOR pathway
MiR-199a-modified exosomes from adipose tissue-derived mesenchymal stem cells improve hepatocellular carcinoma chemosensitivity through mTOR pathway
复制标题
来自脂肪组织来源的间充质干细胞的 MiR-199a 修饰的外泌体通过 mTOR 途径改善肝细胞癌的化疗敏感性
DOI：
10.1186/s13046-019-1512-5
复制
发表时间：
2020-01-02
影响因子：
11.3
通讯作者：
Liu, Yanning
中科院分区：
文献类型：
--
作者：
Lou, Guohua;Chen, Liang;Liu, Yanning
背景miR-199a-3p(miR-199a)可增强肝细胞癌的化疗敏感性。由于miRNA容易通过直接输注降解，有效的载体介导的miR-199a可能是改进肝癌化疗的一种新策略。考虑到间充质干细胞(MSC)来源的外切体是药物和分子转运的天然纳米载体，我们的目的是确定脂肪组织来源的MSCs(AMSCs)是否可以用来运送miR-199a，并提高肝癌的化疗敏感性。方法通过miR-199a慢病毒感染和嘌呤霉素筛选，构建miR-199a修饰的AMSC。从AMSC-199a的培养上清液中分离到MIR-199修饰的外切体(AMSC-Exo-199a)，并用透射电子显微镜、纳米颗粒跟踪分析和流式细胞仪分析对其进行了鉴定。用实时荧光定量聚合酶链式反应检测miR-199a在肝细胞癌标本、AMSCs、外切体和肝癌细胞中的表达水平。用细胞增殖和细胞凋亡率和静脉注射法检测AMSC-Exo-199a对肝癌化疗敏感性的影响。注射入阿霉素治疗的小鼠原位肝癌模型。免疫印迹法检测肝癌细胞和组织中mTOR、p-4EBP1和p-70S6K的表达水平。结果AMSC-Exo-199a具有典型的外切体特性，能有效地介导miR-199a对肝癌细胞的转运。此外，AMSC-Exo-199a通过靶向mTOR并随后抑制mTOR通路而显著增敏肝癌细胞对阿霉素的敏感性。此外，静脉注射AMSC-Exo-199a可分布于肿瘤组织，显著增强Dox的体内抗肝癌作用。结论AMSC-Exo-199a可作为miR-199a的有效载体，通过靶向mTOR通路有效地增敏化疗药物。AMSC-Exo-199a给药可能为提高肝癌的化疗敏感性提供一种新的策略。
Background MiR-199a-3p (miR-199a) can enhance the chemosensitivity of hepatocellular carcinoma (HCC). Because of the easy degradation of miRNA by direct infusion, effective vehicle-mediated delivery of miR-199a may represent a new strategy for improving HCC chemotherapy. Considering mesenchymal stem cell (MSC)-derived exosomes as promising natural nanovectors for drug and molecule delivery, we aimed to determine whether exosomes from adipose tissue-derived MSCs (AMSCs) could be used to deliver miR-199a and improve HCC chemosensitivity. Methods MiR-199a-modified AMSCs (AMSC-199a) were constructed by miR-199a lentivirus infection and puromycin selection. MiR-199-modified exosomes (AMSC-Exo-199a) were isolated from the supernatant of AMSC-199a and were assessed by transmission electron microscopy, nanoparticle tracking analysis, and flow cytometry analysis. The expression levels of miR-199a in HCC samples, AMSCs, exosomes, and HCC cells were quantified by real-time PCR. The effects of AMSC-Exo-199a on HCC chemosensitivity were determined by cell proliferation and apoptosis assays and by i.v. injection into orthotopic HCC mouse models with doxorubicin treatment. MTOR, p-4EBP1 and p-70S6K levels in HCC cells and tissues were quantified by Western blot. Results AMSC-Exo-199a had the classic characteristics of exosomes and could effectively mediate miR-199a delivery to HCC cells. Additionally, AMSC-Exo-199a significantly sensitized HCC cells to doxorubicin by targeting mTOR and subsequently inhibiting the mTOR pathway. Moreover, i.v.-injected AMSC-Exo-199a could distribute to tumor tissue and markedly increased the effect of Dox against HCC in vivo. Conclusions AMSC-Exo-199a can be an effective vehicle for miR-199a delivery, and they effectively sensitized HCC to chemotherapeutic agents by targeting mTOR pathway. AMSC-Exo-199a administration may provide a new strategy for improving HCC chemosensitivity.