Induction of artificial cancer stem cells from tongue cancer cells by defined reprogramming factors.
Induction of artificial cancer stem cells from tongue cancer cells by defined reprogramming factors.
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DOI：
10.1186/s12885-016-2416-9
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发表时间：
2016-07-27
期刊：
影响因子：
3.8
通讯作者：
Ueyama Y
中科院分区：
文献类型：
--
作者：
Harada K;Ferdous T;Cui D;Kuramitsu Y;Matsumoto T;Ikeda E;Okano H;Ueyama Y
肿瘤干细胞（cancer stem cells，CSCs）是肿瘤中的一小部分细胞亚群，与肿瘤的发生、生长、复发和转移以及肿瘤对药物和放疗的抵抗有关。CSC是开发癌症治疗新策略的重要靶标。然而，由于缺乏简单可靠的方法来分离、收集和维持足够数量的CSC，目前以CSC为靶点的新抗癌药物的发现受到阻碍。在这里，我们研究了将定义的重编程因子（Oct 4、shp 53、Sox 2、Klf 4、l-Myc和Lin 28）引入HSC 2舌癌细胞中是否可以将HSC 2转化为具有CSC特性的HSC 2。我们通过电穿孔的方法将定义的重编程因子通过附加型载体导入HSC 2舌癌细胞中以产生转染细胞。我们通过体外细胞增殖实验、迁移实验、伤口愈合实验、成球实验、化疗敏感性和放射敏感性实验研究了转染细胞的恶性特性，并检测了转染细胞的体内致瘤潜力。转染细胞（HSC 2/hOCT 3/4-shp 53-F、HSC 2/hSK、HSC 2/hUL、HSC 2/hOCT 3/4-shp 53-F + hSK、HSC 2/hOCT 3/4-shp 53-F + hUL、HSC 2/hSK + hUL、HSC 2/hOCT 3/4-shp 53-F + hSK + hUL）在培养中显示恶性表型，并且比亲本HSC 2和对照HSC 2/EGFP转染细胞更有效地在裸鼠背部形成肿瘤。与HSC 2和HSC 2/EGFP相比，它们表现出对化疗药物的耐药性增加; 5-氟尿嘧啶、顺铂、多西他赛、三氟胸苷、唑来膦酸、西妥昔单抗、硼替佐米和放射。在所有转染的细胞中，含有所有重编程因子的HSC 2/hOCT 3/4-shp 53-F + hSK + hUL细胞显示出最具侵袭性和恶性性质，并且在含有人重组成纤维细胞生长因子-2（FGF-2）和表皮生长因子（EGF）的培养基中呈现出最高的球体数目。这些发现表明，通过诱导细胞重编程获得的人工肿瘤干细胞可能有助于研究潜在恶性肿瘤的获得以及筛选CSCs靶向药物。
The cancer stem cells (CSCs), a small subpopulation of cells in tumor are responsible for the tumor initiation, growth, recurrence and metastasis of cancer, as well as resistance of cancers to drugs or radiotherapy. CSCs are an important target for the development of novel strategies in cancer treatment. However, CSCs-targeted new anti-cancer drug discovery is currently hindered by the lack of easy and reliable methods for isolating, collecting and maintaining sufficient number of CSCs. Here, we examined whether introduction of defined reprogramming factors (Oct4, shp53, Sox2, Klf4, l-Myc and Lin28) into HSC2 tongue cancer cells could transform the HSC2 into HSC2 with CSCs properties. We introduced the defined reprogramming factors into HSC2 tongue cancer cells via episomal vectors by electroporation method to generate transfectant cells. We investigated the malignant properties of the transfectant cells by cell proliferation assay, migration assay, wound healing assay, sphere formation assay, chemosensitivity and radiosensitivity assay in vitro; and also examined the tumorigenic potential of the transfectants in vivo. The transfectant cells (HSC2/hOCT3/4-shp53-F, HSC2/hSK, HSC2/hUL, HSC2/hOCT3/4-shp53-F + hSK, HSC2/hOCT3/4-shp53-F + hUL, HSC2/hSK + hUL, HSC2/hOCT3/4-shp53-F + hSK + hUL) displayed a malignant phenotype in culture and form tumors on the back of nude mice more efficiently than parental HSC2 and control HSC2/EGFP transfectant cells. They exhibited increased resistance to chemotherapeutic agents; 5-fluorouracil, cisplatin, docetaxel, trifluorothymidine, zoledronic acid, cetuximab, bortezomib and radiation when compared with HSC2 and HSC2/EGFP. Among all the transfected cells, HSC2/hOCT3/4-shp53-F + hSK + hUL cell containing all of the reprogramming factors showed the most aggressive and malignant properties and presented the highest number of spheres in the culture medium containing human recombinant fibroblast Growth Factor-2 (FGF-2) and epidermal Growth Factor (EGF). These findings suggest that artificial cancer stem cells obtained by the induction of cellular reprogramming may be useful for investigating the acquisition of potential malignancy as well as screening the CSCs-targeting drugs.