Identification of Serum miR-501-3p and miR-338-3p as Novel Diagnostic Biomarkers for Breast Cancer and Their Target Genes Associated with Immune Infiltration

Abstract

Background

MicroRNAs influence the growth and metastasis of breast cancer (BC) by regulating their target genes. Our study aims to screen and identify miRNAs that are closely related to the development of breast cancer, and explore the role of these miRNAs and their target genes in breast cancer.

Methods

Bioinformatics tools were applied to screen breast cancer-associated miRNAs and predict their potential target genes. Serum miRNAs were measured using RT-PCR. The correlation between miRNA expression and different clinicopathological features of BC patients was analyzed. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value. GEPIA, Kaplan-Meier Plotter, TIMER, and TISIDB databases were used to validate the expression levels and their prognostic value, as well as their target gene associated with immune infiltrating cells and immune checkpoints.

Results

Breast cancer-associated serum miR-338-3p and miR-501-3p were screened and verified for the first time. Serum miR-501-3p was elevated in BC and was closely linked to the ki-67 index and histological grade. CDKN2C, as a potential target gene of miR-501-3p, was enriched in the cGMP-PKG signaling pathway. Serum miR-338-3p was reduced in BC and was strongly linked to lymph node metastasis and histological grading. ACTR2, CDH1, COL1A1, RBBP5, RRM1, and TPM3, as potential target genes of miR-338-3p, were enriched in MAPK, PI3K-Akt, and RAS signaling pathways. These target genes were found to be linked to breast cancer prognosis, immune infiltrating cells, and immune checkpoint inhibitors. Analysis of ROC curve showed that serum miR-501-3p combined with serum miR-338-3p had a high diagnostic value in breast cancer (AUC: 0.89, 95% CI: 0.821–0.958).

Conclusion

Serum miR-501-3p combined with serum miR-338-3p show obvious clinical significance in the diagnosis and prognosis of breast cancer, which suggests that they may act as novel diagnostic biomarkers for breast cancer.

Keywords:

• breast cancer
• miR-501-3p
• miR-338-3p
• serum biomarker
• bioinformatics analysis

Abbreviations

BC, Breast cancer; DEMs, Differentially Expressed MicroRNAs; ROC, Receiver operating characteristic curve; AUC, The area under the Curve; GO, Gene Ontology; KEGG, Kyoto Encyclopaedia of Genes and Genomes; PPI, Protein-Protein Interaction network.

Data Sharing Statement

All data during the course of this study are included in this article.

Ethics Approval

This study design was approved by the Medical Ethics Committee of Weifang People’s Hospital of Shandong Province (KYLL20230120-2) and all patients provided informed consent, in accordance with the Declaration of Helsinki.

Consent to Participate

Obtain informed consent from all participants.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

The authors declare no competing interests.

Additional information

Funding

The present work was supported by the Youth Natural Science Foundation of Shandong Province (ZR2021QH102), the Science and Technology Development Plan Project of Weifang City (2021YX040), the Students’ Research Fund of Weifang Medical University (X2021615, X2021641, X2021186).