利用人黑色素瘤细胞裂解液建立人酪氨酸酶活性抑制试验。

IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS BioTechniques Pub Date : 2024-11-01 Epub Date: 2024-12-16 DOI:10.1080/07366205.2024.2441637
Gengxuan Shi, Yunjiang Feng, Kathryn F Tonissen
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引用次数: 0

摘要

治疗皮肤色素沉着需要酪氨酸酶(TYR)抑制剂。目前,活细胞 TYR 试验和蘑菇 TYR 体外试验是筛选 TYR 抑制剂的常用方法。然而,这些方法要么耗时费钱,要么不具有人类 TYR(hsTYR)特异性。在此,我们介绍了一种简单的 hsTYR 检测方法,该方法使用色素人类黑色素瘤细胞系制备的细胞裂解物,收集细胞颗粒后只需不到 3 小时即可完成。我们使用已知的 hsTYR 抑制剂曲酸作为阳性对照,证实了该检测方法具有物种特异性,而抑制蘑菇 TYR 但不抑制 hsTYR 的熊果苷则不起作用。在活体生物模型中进行后续研究之前,这种测定是一种确认 hsTYR 抑制作用的简单方法。
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Development of a human tyrosinase activity inhibition assay using human melanoma cell lysate.

Tyrosinase (TYR) inhibitors are required to treat skin hyperpigmentation. Currently live cell-based TYR assays and mushroom TYR in vitro assays are the common methods used to screen for TYR inhibitors. However, these methods are either time consuming and expensive or are not human TYR (hsTYR) specific. Here, we describe a simple hsTYR assay using cell lysate prepared from pigmented human melanoma cell lines that takes less than 3 hours to complete after collecting cell pellets. We confirmed the assay is species specific by using a known hsTYR inhibitor, kojic acid, as a positive control, while arbutin, which inhibits mushroom TYR, but not hsTYR, was not effective. This assay is a simple method to confirm hsTYR inhibition before conducting follow-up studies in live biological models.

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来源期刊
BioTechniques
BioTechniques 工程技术-生化研究方法
CiteScore
4.40
自引率
0.00%
发文量
68
审稿时长
3.3 months
期刊介绍: BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.
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