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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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.