Elastase Reduces Background Autofluorescence in ALK Fluorescence In Situ Hybridization Assays for Lung Cancers.

IF 4.1 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biomedical Journal Pub Date : 2025-03-06 DOI:10.1016/j.bj.2025.100840

Sheng-Chi Hsu, Tsai-Hsien Hung, Hsiao-Chun Wu, Kwai-Fong Ng, Tse-Ching Chen

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引用次数: 0

Abstract

Background: Anaplastic lymphoma kinase (ALK) inhibitors have been effective in treating non-small cell lung cancers (NSCLC) with ALK translocation. However, high background autofluorescence in lung tissues interferes with fluorescence in situ hybridization (FISH) assays, masking molecular probe signals and hindering data interpretation.

Materials and methods: To reduce autofluorescence, NSCLC tissue sections were treated with various proteases, including collagenase types I, II, IV, and elastase, to determine the most effective enzyme. We then conducted ALK break-apart FISH assays on 120 NSCLC samples comparing standard and novel pretreatment protocols.

Results: Elastase was identified as the most effective enzyme for reducing autofluorescence while preserving nuclear integrity. The elastase-based pretreatment enabled clear FISH signal detection in all cases, reducing the retest rate from 86.7% to 0%. Furthermore, two additional ALK translocated cases were detected with elastase pretreatment, which were indeterminable with pepsin treatment alone.

Conclusions: This novel elastase pretreatment protocol addresses autofluorescence interference in lung tissues and can significantly improve the reliability of FISH assays for targeted therapy decisions.

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弹性酶在肺癌的ALK荧光原位杂交检测中降低了背景自身荧光。

背景：间变性淋巴瘤激酶（ALK）抑制剂对ALK易位的非小细胞肺癌（NSCLC）有效。然而，肺组织中的高背景自身荧光干扰荧光原位杂交（FISH）检测，掩盖分子探针信号并阻碍数据解释。材料和方法：为了减少自身荧光，用各种蛋白酶处理NSCLC组织切片，包括胶原酶I、II、IV和弹性酶，以确定最有效的酶。然后，我们对120个NSCLC样本进行了ALK分解FISH测定，比较了标准和新型预处理方案。结果：Elastase是在保持核完整性的同时减少自身荧光最有效的酶。基于弹性酶的预处理在所有情况下都能检测到清晰的FISH信号，将复测率从86.7%降低到0%。此外，另外两例ALK易位病例是用弹性蛋白酶预处理检测到的，单独用胃蛋白酶治疗是不确定的。结论：这种新的弹性蛋白酶预处理方案解决了肺组织中的自身荧光干扰，可以显著提高FISH检测在靶向治疗决策中的可靠性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biomedical Journal Medicine-General Medicine

CiteScore

11.60

自引率

1.80%

发文量

128

审稿时长

42 days

期刊介绍： Biomedical Journal publishes 6 peer-reviewed issues per year in all fields of clinical and biomedical sciences for an internationally diverse authorship. Unlike most open access journals, which are free to readers but not authors, Biomedical Journal does not charge for subscription, submission, processing or publication of manuscripts, nor for color reproduction of photographs. Clinical studies, accounts of clinical trials, biomarker studies, and characterization of human pathogens are within the scope of the journal, as well as basic studies in model species such as Escherichia coli, Caenorhabditis elegans, Drosophila melanogaster, and Mus musculus revealing the function of molecules, cells, and tissues relevant for human health. However, articles on other species can be published if they contribute to our understanding of basic mechanisms of biology. A highly-cited international editorial board assures timely publication of manuscripts. Reviews on recent progress in biomedical sciences are commissioned by the editors.