Interferon-β gene-modified human bone marrow mesenchymal stem cells attenuate hepatocellular carcinoma through inhibiting AKT/FOXO3a pathway.
Interferon-β gene-modified human bone marrow mesenchymal stem cells attenuate hepatocellular carcinoma through inhibiting AKT/FOXO3a pathway.
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干扰素-β基因改性的人骨骨髓间充质干细胞通过抑制AKT/FOXO3A途径减弱肝细胞癌。
DOI：
10.1038/bjc.2013.422
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发表时间：
2013-09-03
影响因子：
8.8
通讯作者：
Gao ZL
中科院分区：
文献类型：
--
作者：
Xie C;Xie DY;Lin BL;Zhang GL;Wang PP;Peng L;Gao ZL
本研究旨在探讨基因工程诱导的骨髓间充质干细胞（BMSCs）作为基因递送系统治疗肝细胞癌（HCC）的体内外研究。为了测量对体外肿瘤细胞生长的影响，将产生IFN-β的BMSC（BMSC/IFN-β）与HCC细胞系HepG 2和Huh 7共培养。采用酶联免疫吸附试验（ELISA）检测BMSC条件培养液（CM）中IFN-β的分泌。采用MTT法、克隆形成实验、BrdU染色、细胞周期分析和NOD/SCID小鼠移植瘤模型，观察BMSC/IFN-β对肝癌细胞增殖的影响。为了检查BMSC/IFN-β对AKT/FOXO 3a信号传导的影响，进行RT-PCR和蛋白质印迹。BMSC/IFN-β细胞能稳定分泌高水平的IFN-β。MTT和集落形成实验均显示，BMSC/IFN-β-CM培养的肝癌细胞生长速度低于BMSC/vector-CM或DMEM培养组。与BMSC/IFN-β-CM共培养显著降低了掺入BrdUrd的细胞百分比。BMSC/IFN-β-CM处理后，G1期细胞比例增加，S期细胞比例减少。BMSC/IFN-β对NOD/SCID小鼠肝癌的生长有抑制作用，并延长了小鼠的生存期。与对照组相比，BMSC/IFN-β-CM共培养的肝癌细胞p21、p27表达增加，cyclin D1和Rb磷酸化表达降低。它与Akt活性的抑制和FOXO 3a转录活性的增强有关。IFN-β基因修饰的BMSC可以通过抑制AKT/FOXO 3a途径有效抑制肝癌细胞的体内外增殖。这些结果表明BMSC/IFN-β是一种有效的肝癌细胞治疗工具。
This study aims to investigate the using of bone marrow mesenchymal stem cells (BMSCs) genetically engineered to produce interferon-β (IFN-β) as a gene delivery system to treat hepatocellular carcinoma (HCC) in vitro and in vivo. To measure the effects on tumour cell growth in vitro, IFN-β-producing BMSCs (BMSC/IFN-β) were co-cultured with the HCC cell line HepG2 and Huh7. Enzyme-linked immunosorbent assay (ELISA) was used to detect the IFN-β secretion in the BMSC culture condition medium (CM). The effect of BMSC/IFN-β on HCC cells proliferation was examined both in vitro and in vivo by using MTT, colony formation assay, BrdU staining, cell cycle analysis, and xenografted NOD/SCID mouse tumour model. To examine the impact of BMSC/IFN-β on the AKT/FOXO3a signalling, RT–PCR and western blotting were performed. The BMSC/IFN-β cells can stably secrete high levels of IFN-β. Both MTT and colony forming assay showed that HCC cells had a lower growth rate when cultured in BMSC/IFN-β-CM as compared with that in BMSC/vector-CM or DMEM culture group. Co-culture with BMSC/IFN-β-CM dramatically decreased the percentages of cells with incorporated BrdUrd. In BMSC/IFN-β-CM-treated HCC cells, the proportion of G1-phase cells increased but it decreased in the S phase of the cell. The BMSC/IFN-β inhibited HCC growth in NOD/SCID mice and proved the survival period of these mice. Compared with the control group, p21 and p27 expression of hepatoma cells increased, whereas cyclin D1 and phosphorylation of Rb expression decreased when co-cultured with BMSC/IFN-β-CM. It was associated with suppression of Akt activity and enhanced transcriptional activity of FOXO3a. The IFN-β gene-modified BMSCs can effectively inhibit the proliferation of HCC cells in vitro and in vivo through inhibiting AKT/FOXO3a pathway. These results indicate that BMSC/IFN-β are a powerful anticancer cytotherapeutic tool for HCC.
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影响因子：
11.2
作者：
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通讯作者：
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