Wu Zheng, Liu Wei, Wang Zujia, Zeng Baitao, Peng Guangnan, Niu Hongyan, Chen Linlin, Liu Cong, Hu Qian, Zhang Yuxuan, Pan Mengmeng, Wu Lingqian, Liu Mujun, Liu Xionghao, Liang Desheng
1Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha, Hunan China.
3Hunan Key Laboratory of Animal Model for Human Diseases, Central South University, Changsha, Hunan China.
Cancer Cell Int. 2020 Jan 30;20:33. doi: 10.1186/s12935-020-1112-7. eCollection 2020.
Interleukin-24 (-) is a therapeutic gene for melanoma, which can induce melanoma cell apoptosis. Mesenchymal stem cells (MSCs) show promise as a carrier to delivery anti-cancer factors to tumor tissues. Induced pluripotent stem cells (iPSCs) are an alternative source of mesenchymal stem cells (MSCs). We previously developed a novel non-viral gene targeting vector to target - to human iPSCs. This study aims to investigate whether MSCs derived from the iPSCs with the site-specific integration of - can inhibit the growth of melanoma in a tumor-bearing mouse model via retro-orbital injection.
IL-24-iPSCs were differentiated into IL-24-iMSCs in vitro, of which cellular properties and potential of differentiation were characterized. The expression of IL-24 in the IL-24-iMSCs was measured by qRT-PCR, Western Blotting, and ELISA analysis. IL-24-iMSCs were transplanted into the melanoma-bearing mice by retro-orbital intravenous injection. The inhibitory effect of IL-24-iMSCs on the melanoma cells was investigated in a co-culture system and tumor-bearing mice. The molecular mechanisms underlying IL-24-iMSCs in exerting anti-tumor effect were also explored.
iPSCs-derived iMSCs have the typical profile of cell surface markers of MSCs and have the ability to differentiate into osteoblasts, adipocytes, and chondroblasts. The expression level of IL-24 in IL-24-iMSCs reached 95.39 ng/10 cells/24 h, which is significantly higher than that in iMSCs, inducing melanoma cells apoptosis more effectively in vitro compared with iMSCs. IL-24-iMSCs exerted a significant inhibitory effect on the growth of melanoma in subcutaneous mouse models, in which the migration of IL-24-iMSCs to tumor tissue was confirmed. Additionally, increased expression of Bax and Cleaved caspase-3 and down-regulation of Bcl-2 were observed in the mice treated with IL-24-iMSCs.
MSCs derived from iPSCs with the integration of - at rDNA locus can inhibit the growth of melanoma in tumor-bearing mouse models when administrated via retro-orbital injection.
白细胞介素-24（IL-24）是一种用于黑色素瘤的治疗性基因，可诱导黑色素瘤细胞凋亡。间充质干细胞（MSCs）有望作为将抗癌因子递送至肿瘤组织的载体。诱导多能干细胞（iPSCs）是间充质干细胞的一种替代来源。我们之前开发了一种新型非病毒基因靶向载体，用于靶向人iPSCs。本研究旨在探讨经位点特异性整合IL-24的iPSCs来源的MSCs能否通过眶后注射在荷瘤小鼠模型中抑制黑色素瘤的生长。
将IL-24-iPSCs体外分化为IL-24-iMSCs，并对其细胞特性和分化潜能进行表征。通过qRT-PCR、蛋白质免疫印迹和ELISA分析检测IL-24-iMSCs中IL-24的表达。将IL-24-iMSCs通过眶后静脉注射移植到荷黑色素瘤小鼠体内。在共培养系统和荷瘤小鼠中研究IL-24-iMSCs对黑色素瘤细胞的抑制作用。还探讨了IL-24-iMSCs发挥抗肿瘤作用的分子机制。
iPSCs来源的iMSCs具有MSCs典型的细胞表面标志物特征，并且具有分化为成骨细胞、脂肪细胞和软骨细胞的能力。IL-24-iMSCs中IL-24的表达水平达到95.39 ng/10⁶细胞/24 h，显著高于iMSCs，与iMSCs相比，在体外能更有效地诱导黑色素瘤细胞凋亡。IL-24-iMSCs对皮下小鼠模型中的黑色素瘤生长具有显著抑制作用，证实了IL-24-iMSCs向肿瘤组织的迁移。此外，在用IL-24-iMSCs处理的小鼠中观察到Bax和Cleaved caspase-3表达增加以及Bcl-2表达下调。
经rDNA位点整合IL-24的iPSCs来源的MSCs通过眶后注射给药时，可在荷瘤小鼠模型中抑制黑色素瘤的生长。
