Laboratory of Veterinary Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, 361-763, Republic of Korea.
Mol Oncol. 2013 Jun;7(3):543-54. doi: 10.1016/j.molonc.2013.01.004. Epub 2013 Jan 19.
Genetically engineered stem cells (GESTECs) exhibit a potent therapeutic efficacy via their strong tumor tropism toward cancer cells. In this study, we introduced the human parental neural stem cells, HB1.F3, with the human interferon beta (IFN-β) gene which is a typical cytokine gene that has an antitumor effect and the cytosine deaminase (CD) gene from Escherichia coli (E. coli) that could convert the non-toxic prodrug, 5-fluorocytosine (5-FC), to a toxic metabolite, 5-fluorouracil (5-FU). Two types of stem cells expressing the CD gene (HB1.F3.CD cells) and both the CD and human IFN-β genes (HB1.F3.CD.IFN-β) were generated. The present study was performed to examine the migratory and therapeutic effects of these GESTECs against the colorectal cancer cell line, HT-29. When co-cultured with colorectal cancer cells in the presence of 5-FC, HB1.F3.CD and HB1.F3.CD.IFN-β cells exhibited the cytotoxicity on HT-29 cells via the bystander effect. In particular, HB1.F3.CD.IFN-β cells showed the synergistic cytotoxic activity of 5-FU and IFN-β. We also confirmed the migration ability of HB1.F3.CD and HB1.F3.CD.IFN-β cells toward HT-29 cells by a modified migration assay in vitro, where chemoattractant factors secreted by HT-29 cells attracted the GESTECs. In a xenograft mouse model, the volume of tumor mass was decreased up to 56% in HB1.F3.CD injected mice while the tumor mass was greatly inhibited about 76% in HB1.F3.CD.IFN-β injected mice. The therapeutic treatment by these GESTECs is a novel strategy where the combination of the migration capacity of stem cells as a vector for therapeutic genes towards colorectal cancer and a synergistic antitumor effect of CD and IFN-β genes can selectively target this type of cancer.
基因工程化的干细胞(GESTECs)通过其对癌细胞的强烈肿瘤趋向性表现出强大的治疗功效。在这项研究中,我们引入了人源亲本神经干细胞 HB1.F3,其中包含人干扰素β(IFN-β)基因,这是一种具有抗肿瘤作用的典型细胞因子基因,以及来自大肠杆菌(E. coli)的胞嘧啶脱氨酶(CD)基因,它可以将非毒性前体药物 5-氟胞嘧啶(5-FC)转化为有毒代谢物 5-氟尿嘧啶(5-FU)。生成了两种表达 CD 基因的干细胞(HB1.F3.CD 细胞)和同时表达 CD 和人 IFN-β 基因的两种干细胞(HB1.F3.CD.IFN-β)。本研究旨在检验这些 GESTECs 对结直肠癌细胞系 HT-29 的迁移和治疗作用。当与结直肠癌细胞在 5-FC 存在下共培养时,HB1.F3.CD 和 HB1.F3.CD.IFN-β 细胞通过旁观者效应对 HT-29 细胞表现出细胞毒性。特别是,HB1.F3.CD.IFN-β 细胞表现出 5-FU 和 IFN-β 的协同细胞毒性活性。我们还通过体外改良迁移测定法证实了 HB1.F3.CD 和 HB1.F3.CD.IFN-β 细胞向 HT-29 细胞的迁移能力,其中 HT-29 细胞分泌的趋化因子吸引 GESTECs。在异种移植小鼠模型中,HB1.F3.CD 注射小鼠的肿瘤体积减少了 56%,而 HB1.F3.CD.IFN-β 注射小鼠的肿瘤体积则大大抑制了约 76%。这些 GESTECs 的治疗方法是一种新策略,其中干细胞的迁移能力作为治疗基因向结直肠癌细胞的载体与 CD 和 IFN-β 基因的协同抗肿瘤作用相结合,可以选择性地针对这种类型的癌症。