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人口腔鳞状细胞癌(OSCC)H103和H376细胞系对逆转录病毒OSKM介导的重编程的敏感性。

Susceptibility of Human Oral Squamous Cell Carcinoma (OSCC) H103 and H376 cell lines to Retroviral OSKM mediated reprogramming.

作者信息

Verusingam Nalini Devi, Yeap Swee Keong, Ky Huynh, Paterson Ian C, Khoo Suan Phaik, Cheong Soon Keng, Ong Alan H K, Kamarul Tunku

机构信息

Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Selangor, Malaysia.

Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.

出版信息

PeerJ. 2017 Apr 13;5:e3174. doi: 10.7717/peerj.3174. eCollection 2017.

Abstract

Although numbers of cancer cell lines have been shown to be successfully reprogrammed into induced pluripotent stem cells (iPSCs), reprogramming Oral Squamous Cell Carcinoma (OSCC) to pluripotency in relation to its cancer cell type and the expression pattern of pluripotent genes under later passage remain unexplored. In our study, we reprogrammed and characterised H103 and H376 oral squamous carcinoma cells using retroviral OSKM mediated method. Reprogrammed cells were characterized for their embryonic stem cells (ESCs) like morphology, pluripotent gene expression via quantitative real-time polymerase chain reaction (RT-qPCR), immunofluorescence staining, embryoid bodies (EB) formation and directed differentiation capacity. Reprogrammed H103 (Rep-H103) exhibited similar ESCs morphologies with flatten cells and clear borders on feeder layer. Reprogrammed H376 (Rep-H376) did not show ESCs morphologies but grow with a disorganized morphology. Critical pluripotency genes Oct4, Sox2 and Nanog were expressed higher in Rep-H103 against the parental counterpart from passage 5 to passage 10. As for Rep-H376, Nanog expression against its parental counterpart showed a significant decrease at passage 5 and although increased in passage 10, the level of expression was similar to the parental cells. Rep-H103 exhibited pluripotent signals (Oct4, Sox2, Nanog and Tra-1-60) and could form EB with the presence of three germ layers markers. Rep-H103 displayed differentiation capacity into adipocytes and osteocytes. The OSCC cell line H103 which was able to be reprogrammed into an iPSC like state showed high expression of Oct4, Sox2 and Nanog at late passage and may provide a potential iPSC model to study multi-stage oncogenesis in OSCC.

摘要

尽管已证明许多癌细胞系能够成功重编程为诱导多能干细胞(iPSC),但关于口腔鳞状细胞癌(OSCC)如何根据其癌细胞类型重编程为多能性以及传代后期多能基因的表达模式仍未得到探索。在我们的研究中,我们使用逆转录病毒OSKM介导的方法对H103和H376口腔鳞状癌细胞进行了重编程和表征。对重编程细胞的胚胎干细胞(ESC)样形态、通过定量实时聚合酶链反应(RT-qPCR)检测多能基因表达、免疫荧光染色、胚状体(EB)形成和定向分化能力进行了表征。重编程的H103(Rep-H103)在饲养层上表现出与ESC相似的形态,细胞扁平且边界清晰。重编程的H376(Rep-H376)未表现出ESC形态,而是以无序形态生长。关键多能性基因Oct4、Sox2和Nanog在Rep-H103中从第5代到第10代相对于亲代细胞表达更高。至于Rep-H376,Nanog相对于亲代细胞的表达在第5代时显著降低,尽管在第10代有所增加,但表达水平与亲代细胞相似。Rep-H103表现出多能信号(Oct4、Sox2、Nanog和Tra-1-60)并且可以形成具有三个胚层标志物的EB。Rep-H103表现出向脂肪细胞和骨细胞的分化能力。能够重编程为类似iPSC状态的OSCC细胞系H103在传代后期Oct4、Sox2和Nanog表达较高,可能为研究OSCC的多阶段肿瘤发生提供一个潜在的iPSC模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5eae/5392249/c9f7e8f4ae4c/peerj-05-3174-g001.jpg

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