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开发一种荧光素酶/荧光素细胞增殖(XenoLuc)检测法,用于实时测量共培养系统中 GFP-Luc2 修饰细胞。

Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system.

机构信息

Molecular Pathology Unit, Cancer Research Centre, Institute for Medical Research, National Institutes of Health (NIH Complex), Ministry of Health Malaysia, Level 4, Block C7, No: 1, Jalan Setia Murni U13/52, Section U13, Setia Alam, 40170 Shah Alam, 50588, Kuala Lumpur, Selangor, Malaysia.

Present Address: Department of Medical Sciences, School of Healthcare and Medical Sciences, Sunway University, 47500, Bandar Sunway, Selangor Darul Ehsan, Malaysia.

出版信息

BMC Biotechnol. 2019 Jun 14;19(1):34. doi: 10.1186/s12896-019-0528-4.

Abstract

BACKGROUND

In vitro modelling of cancer cells is becoming more complex due to prevailing evidence of intimate interactions between cancer cells and their surrounding stroma. A co-culture system which consists of more than one cell type is physiologically more relevant and thus, could serve as a useful model for various biological studies. An assay that specifically detects the phenotypic changes of cancer cells in a multi-cellular system is lacking for nasopharyngeal carcinoma (NPC).

RESULTS

Here, we describe a luciferase/luciferin (XenoLuc) assay that could specifically measure changes in the proliferation of cancer cells in the co-culture system using two modified NPC patient-derived tumour xenograft (PDTXs) cells: Xeno284-gfp-luc2 and XenoB110-gfp-luc2. Through this assay, we are able to show that the growth of NPC xenograft cells in both two-dimensional (2D) and three-dimensional (3D) models was enhanced when co-cultured with normal human dermal fibroblasts (NHDFs). In addition, potential applications of this assay in in vitro drug or inhibitor screening experiments are also illustrated.

CONCLUSIONS

XenoLuc assay is specific, sensitive, rapid and cost-effective for measuring the growth of luciferase-expressing cells in a co- or multiple-culture system. This assay may also be adapted for tumour microenvironment studies as well as drug screening experiments in more complex 3D co-culture systems.

摘要

背景

由于越来越多的证据表明癌细胞与其周围基质之间存在密切的相互作用,因此癌细胞的体外建模变得越来越复杂。由多种细胞类型组成的共培养系统在生理上更为相关,因此可以作为各种生物学研究的有用模型。目前尚缺乏用于鼻咽癌(NPC)的特定检测多细胞系统中癌细胞表型变化的分析方法。

结果

在这里,我们描述了一种荧光素酶/荧光素(XenoLuc)分析方法,该方法可以使用两种改良的 NPC 患者来源的肿瘤异种移植(PDTX)细胞:Xeno284-gfp-luc2 和 XenoB110-gfp-luc2 ,专门检测共培养系统中癌细胞增殖的变化。通过该分析方法,我们能够证明当与正常人皮肤成纤维细胞(NHDFs)共培养时,NPC 异种移植物细胞在二维(2D)和三维(3D)模型中的生长均得到增强。此外,还说明了该分析方法在体外药物或抑制剂筛选实验中的潜在应用。

结论

XenoLuc 分析方法对于测量共培养或多培养系统中表达荧光素酶的细胞的生长具有特异性、敏感性、快速和具有成本效益。该分析方法还可以适应肿瘤微环境研究以及更复杂的 3D 共培养系统中的药物筛选实验。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc84/6570829/6f298fe9c46b/12896_2019_528_Fig1_HTML.jpg

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