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用于非小细胞肺癌的新型特异性mRNA杂交检测法。

Novel -specific mRNA hybridization assay for non-small-cell lung carcinoma.

作者信息

Hirai Noriko, Sasaki Takaaki, Okumura Shunsuke, Sado Masatoshi, Akiyama Naoko, Kitada Masahiro, Takei Hidehiro, Ohsaki Yoshinobu

机构信息

Respiratory Center, Asahikawa Medical University, Asahikawa, Hokkaido, Japan.

Department of Pathology, Asahikawa Medical University, Asahikawa, Hokkaido, Japan.

出版信息

Transl Lung Cancer Res. 2020 Apr;9(2):257-268. doi: 10.21037/tlcr.2020.03.04.

DOI:10.21037/tlcr.2020.03.04
PMID:32420065
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7225155/
Abstract

BACKGROUND

A recent technical advance in mRNA hybridization (mRNA-ISH) assays provides simultaneous signal amplification and background suppression with a unique probe design that enables single-molecule visualization. We assessed the utility of the mRNA-ISH assay as a diagnostic tool for detecting anaplastic lymphoma receptor tyrosine kinase () mRNA in non-small-cell lung carcinoma (NSCLC). We compared the mRNA-ISH assay with immunohistochemistry (IHC) and fluorescence hybridization (FISH).

METHODS

The study included 279 surgically resected lung adenocarcinomas and 44 transbronchial-biopsied (TBB) adenocarcinomas. mRNA-ISH was conducted using the RNAscope 2.0 system, which includes pre-designed probes for detecting the tyrosine kinase domain encoded in mRNA. IHC was conducted on all 323 samples using ALK-specific antibodies. mRNA-ISH was performed on 279 surgical samples and 6 TBB samples. Break-apart FISH was used to examine samples that were mRNA-ISH-positive or IHC-positive.

RESULTS

ALK protein expression was detected in 11 of 279 specimens (3.9%). mRNA was also detected with mRNA-ISH in ALK-positive samples, and 9 of the 11 specimens (81%) were also positive for ALK using break-apart FISH. Using the IHC results as a reference, the sensitivity and specificity of mRNA-ISH was 100%. In the TBB cohort, ALK protein expression was observed in 3 of 44 specimens (6.8%), in which mRNA expression was also detected.

CONCLUSIONS

The mRNA-ISH data were highly correlated with the IHC data, and mRNA-ISH detected mRNA expression in every FISH-positive sample. We conclude that mRNA-ISH could serve as an alternative or complementary method for diagnosing rearrangements in NSCLC.

摘要

背景

mRNA杂交（mRNA原位杂交，mRNA-ISH）检测技术的一项最新进展是，通过独特的探针设计实现了信号的同时放大和背景抑制，从而能够进行单分子可视化。我们评估了mRNA-ISH检测作为检测非小细胞肺癌（NSCLC）中间变性淋巴瘤受体酪氨酸激酶（ALK）mRNA的诊断工具的实用性。我们将mRNA-ISH检测与免疫组织化学（IHC）和荧光原位杂交（FISH）进行了比较。

方法

该研究纳入了279例手术切除的肺腺癌和44例经支气管活检（TBB）的腺癌。使用RNAscope 2.0系统进行mRNA-ISH检测，该系统包括用于检测ALK mRNA中编码的酪氨酸激酶结构域的预设计探针。对所有323个样本使用ALK特异性抗体进行IHC检测。对279个手术样本和6个TBB样本进行mRNA-ISH检测。使用分离FISH检测mRNA-ISH阳性或IHC阳性的样本。

结果

在279个标本中有11个（3.9%）检测到ALK蛋白表达。在ALK阳性样本中也通过mRNA-ISH检测到ALK mRNA，并且在11个标本中有9个（81%）使用分离FISH检测ALK也呈阳性。以IHC结果为参考，mRNA-ISH的敏感性和特异性均为100%。在TBB队列中，44个标本中有3个（6.8%）观察到ALK蛋白表达，其中也检测到ALK mRNA表达。

结论

ALK mRNA-ISH数据与IHC数据高度相关，并且mRNA-ISH在每个FISH阳性样本中均检测到ALK mRNA表达。我们得出结论，mRNA-ISH可作为诊断NSCLC中ALK重排的替代或补充方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/b51ff52b3b47/tlcr-09-02-257-f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/07e34df032ac/tlcr-09-02-257-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/16848b63b06f/tlcr-09-02-257-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/94c6677d288a/tlcr-09-02-257-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/af3a5b3e4d07/tlcr-09-02-257-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/7e17b7eab5c3/tlcr-09-02-257-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/17452179c126/tlcr-09-02-257-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/8f2c203a8b39/tlcr-09-02-257-f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/b51ff52b3b47/tlcr-09-02-257-f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/07e34df032ac/tlcr-09-02-257-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/16848b63b06f/tlcr-09-02-257-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/94c6677d288a/tlcr-09-02-257-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/af3a5b3e4d07/tlcr-09-02-257-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/7e17b7eab5c3/tlcr-09-02-257-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/17452179c126/tlcr-09-02-257-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/8f2c203a8b39/tlcr-09-02-257-f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14d3/7225155/b51ff52b3b47/tlcr-09-02-257-f8.jpg

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