Faculty, Staff and Student Publications

Publication Date

8-2-2025

Journal

Scientific Reports

DOI

10.1038/s41598-025-12780-3

PMID

40753302

PMCID

PMC12318096

PubMedCentral® Posted Date

8-2-2025

PubMedCentral® Full Text Version

Post-print

Abstract

This study aimed to assess the correlation between RNA sequencing (RNA-seq) and immunohistochemistry (IHC) in detecting key cancer biomarkers across solid tumors, and then, to establish RNA-seq thresholds that accurately reflect clinical IHC classifications. Expression levels of nine biomarkers-ESR1, PGR, AR, MKI67, ERBB2, CD274, CDX2, KRT7, and KRT20-were analyzed in 365 formalin-fixed, paraffin-embedded samples from breast, lung, gastrointestinal, and other solid carcinomas. Correlations between RNA-seq data and IHC scores were determined using Spearman's correlation coefficients, with RNA-seq cut-offs established to distinguish positive from negative IHC scores. The results revealed strong correlations for most biomarkers, with coefficients ranging from 0.53 to 0.89. RNA-seq thresholds were confirmed across internal and external cohorts, demonstrating high diagnostic accuracy (up to 98%) and precision in identifying biomarker expression levels. The analysis also highlighted the influence of tumor microenvironment and purity, particularly in the moderate correlation of 0.63 observed for PD-L1. Our study demonstrates that RNA-seq can serve as a robust complementary tool to IHC, offering objective and high-throughput biomarker assessment. The RNA-seq thresholds established provide a reliable method for determining biomarker positivity, supporting the integration of RNA-seq in clinical diagnostics to enhance precision, especially where tumor purity and microenvironment factors are significant.

Keywords

Humans, Biomarkers, Tumor, Immunohistochemistry, Neoplasms, Sequence Analysis, RNA, Tumor Microenvironment, Female, RNA-Seq, Gene Expression Regulation, Neoplastic, Biomarker assessment, Tumor microenvironment, Gene expression correlation, Cancer diagnostics, Immunohistochemistry, RNA sequencing, Cancer microenvironment, Tumour biomarkers, Gene expression analysis, Immunohistochemistry, RNA sequencing

Published Open-Access

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