Human amniotic fluid-derived stem cells expressing cytosine deaminase and thymidine kinase inhibits the growth of breast cancer cells in cellular and xenograft mouse models
Human amniotic fluid-derived stem cells expressing cytosine deaminase and thymidine kinase inhibits the growth of breast cancer cells in cellular and xenograft mouse models
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DOI：
10.1038/cgt.2012.15
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发表时间：
2012-06-01
影响因子：
6.4
通讯作者：
Choi, K-C
中科院分区：
文献类型：
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作者：
Kang, N-H;Hwang, K-A;Choi, K-C
人羊水来源干细胞(HAFSCs)具有多向分化、广泛自我更新和肿瘤靶向性等特点，有望在临床抗癌治疗中发挥重要作用。在这项研究中，我们使用hAFSCs作为靶向向乳腺癌细胞输送治疗性自杀基因的载体。利用hAFSCs产生AF2.CD-TK细胞，以表达编码细菌胞嘧啶脱氨酶(CD)和单纯疱疹病毒胸苷激酶(HSV-TK)的两种自杀基因，这两种自杀基因分别将无毒的前药5-氟胞嘧啶(5-FC)和单磷酸更昔洛韦(GCV-MP)转化为细胞毒性代谢产物5-氟尿嘧啶(5-FU)和更昔洛韦(GCV-TP)。在体外细胞活力实验中，AF2.CD-TK细胞在5-FC、GCV前药或两者联合作用下，对人乳腺癌细胞的生长有抑制作用。当5-FC和GCV混合作用时，细胞存活率出现相加的细胞毒效应。在通过注射MDA-MB-231建立的雌性BALB/c裸鼠移植瘤的动物实验中，AF2 CD-TK细胞在5-FC和GCV的存在下处理显著减少了肿瘤的体积和重量，与5-FU处理的小鼠相同。组织病理学和荧光染色进一步证实AF2.CD-TK细胞定位于肿瘤形成部位。此外，经AF2.CD-TK细胞和两种前药处理的小鼠乳腺组织仍保持其正常结构(如表皮和网状层)，而5-FU处理的小鼠的乳腺组织结构几乎被该药物强大的细胞毒性破坏。综上所述，这些结果表明，AF2.CD-TK细胞可以作为一种新的基于细胞的基因导向的前药系统的优秀载体，以选择性地靶向乳腺恶性肿瘤。
As human amniotic fluid-derived stem cells (hAFSCs) are capable of multiple lineage differentiation, extensive self-renewal and tumor targeting, they may be valuable for clinical anticancer therapies. In this study, we used hAFSCs as vehicles for targeted delivery of therapeutic suicide genes to breast cancer cells. hAFSCs were engineered to produce AF2.CD-TK cells in order to express two suicide genes encoding bacterial cytosine deaminase (CD) and herpes simplex virus thymidine kinase (HSV-TK) that convert non-toxic prodrugs, 5-fluorocytosine (5-FC) and mono-phosphorylate ganciclovir (GCV-MP), into cytotoxic metabolites, 5-fluorouracil (5-FU) and triphosphate ganciclovir (GCV-TP), respectively. In cell viability test in vitro, AF2.CD-TK cells inhibited the growth of MDA-MB-231 human breast cancer cells in the presence of the 5-FC or GCV prodrugs, or a combination of these two reagents. When the mixture of 5-FC and GCV was treated together, an additive cytotoxic effect was observed in the cell viability. In animal experiments using female BALB/c nude mouse xenografts, which developed by injecting MDA-MB-231 cells, treatment with AF2.CD-TK cells in the presence of 5-FC and GCV significantly reduced tumor volume and weight to the same extent seen in the mice treated with 5-FU. Histopathological and fluorescent staining assays further showed that AF2.CD-TK cells were located exactly at the site of tumor formation. Furthermore, breast tissues treated with AF2.CD-TK cells and two prodrugs maintained their normal structures (for example, the epidermis and reticular layers) while breast tissue structures in 5-FU-treated mice were almost destroyed by the potent cytotoxicity of the drug. Taken together, these results indicate that AF2.CD-TK cells can serve as excellent vehicles in a novel therapeutic cell-based gene-directed prodrug system to selectively target breast malignancies.