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Screening and preliminary clinical validation of salivary exosomal mRNA KPNA2 as a novel diagnostic marker for esophageal squamous cell carcinoma

Year: 2025
Type: article
Study design: Observational Study
Abstract: Background Esophageal squamous cell carcinoma (ESCC) is one of the most lethal malignancies worldwide, with a 5-year survival rate of less than 30%. Early diagnosis of ESCC is clinically challenging due to the lack of non-invasive screening and diagnostic methods. Salivary exosomal RNA has been reported to have promising applications in cancer liquid biopsy and in vitro diagnosis. The aim of this study was to investigate the significantly differentially expressed mRNAs in salivary exosomes in ESCC patients and healthy population (HC), so as to provide potential molecular targets for non-invasive diagnosis of ESCC. Methods A total of 90 cases of ESCC patients and healthy volunteers were recruited, saliva and blood samples were collected, exosomes were isolated and total RNA was extracted using the exoRNeasy Midi Kit exosome extraction kit, and salivary exosomes were identified in four samples (ESCC = 2, HC=2) by three methods, namely, transmission electron microscope, particle size analysis and protein blotting. Salivary exosomes, salivary cfRNA, and blood cfRNA (ESCC = 6, HC=6 in each group) were sequenced by mRNA transcriptomics using 12 samples, respectively, and compared with differentially expressed genes (DEGs) identified by transcriptomics sequencing of 24 pairs of ESCC cancers and paracancer tissues under the condition that |log 2 FC| > 1, P < 0.05. Twenty potential candidate biomarkers were screened by taking intersection, expression differential fold analysis and combining with literature reports. Finally, qRT-PCR was used to detect the expression status of the above 20 markers in 40 saliva samples (ESCC = 19, HC=21) for preliminary clinical validation. Findings Firstly, we successfully extracted and identified salivary exosomes, and completed transcriptomics sequencing of salivary exosomes, salivary supernatants, and serum. The results of transcriptome expression comparison with tissues showed that based on this dataset, the salivary exosomes show the most co-upregrated mRNAs (86 upregulated DEGs). Among these 86 DEGs, 20 mRNAs with diagnostic potential were further screened out. qRT-PCR results showed that mRNA KPNA2 was stably expressed in salivary exosomes of ESCC patients and was significantly up-regulated in ESCC patients compared to HC (P < 0.0001) and the area under the curve of the subjects with diagnosed ESCC was 81.70% [95% CI, 66.53% -96.88%] with a sensitivity of 100%, a specificity of 76.91%, and a Youden index of 0.762. Interpretation Salivary exosomal mRNA KPNA2 in ESCC patients showed significantly higher expression compared with the healthy ones, which shows promises in the non-invasive diagnosis of esophageal squamous cell carcinoma.
DOI: 10.1016/j.lanwpc.2024.101421
Language: English
Open Access status: gold
Estimated APC paid: $4,250
OpenAlex ID: W4407631491
FWCI:
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