Upregulation of CircCSPP1, CircNRIP1, and CircSMAD2 in Breast Cancer and Their Potential as Diagnostic Biomarkers

Authors
Mahsa Keshavarz-Fathi 1
Farzaneh Darbeheshti 2
Rangarirai Makuku 1
Parmida Sadat Pezeshki 3
Homa Seyedmirzaei 1
Yaser Mansoori 4
Ali Mohammad Mosadeghrad 5
Nima Rezaei 6
1 School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
2 Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
3 Cancer Immunology Project (CIP), Universal Scientific Education and Research Network (USERN), Tehran, Iran
4 Noncommunicable Disease Research Center, Fasa University of Medical Sciences, Fasa, Iran
5 School of Public Health, Health Information Management Research Center, Tehran University of Medical Sciences, Tehran, Iran
6 Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences, Tehran, Iran
10.61882/ibj.4945
Abstract
Background: Identifying novel diagnostic biomarkers with stable structures, such as circulating circular RNAs (circRNAs) can improve the early detection and management of BC. Herein, we conducted this study to analyze the expression profile of four circRNAs, including circCSPP1 (hsa_circ_0001806), circNRIP1 (hsa_circ_0004771), circSMAD2 (hsa_circ_0000847), and circFOXP1 (hsa_circ_0008234) in BC.

Methods: Tumor tissues and adjacent normal tissues were obtained from patients with sporadic breast cancer (BC). Divergent primers were designed to amplify the target transcripts using quantitative real-time PCR. The expression profiles of circRNAs were analyzed in tumor and adjacent normal tissues. Sanger sequencing was performed to confirm the back-splicing junctions of circRNAs. ROC curves were generated to assess the potential of the mentioned RNAs as diagnostic biomarkers.

Results: We observed a significant upregulation of circCSPP1, circNRIP1, and circSMAD2 in tumor tissue compared to adjacent normal tissues. Among them, circCSPP1 was the most highly upregulated one in tumor samples from 39 patients. Expression of circCSPP1 was significantly higher in estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, human epidermal growth factor receptor-2 (HER2)-negative, and triple-negatives compared to ER-positive, PR-positive, HER2-positive, and non-triple-negative ones. Expression of circNRIP1 was also significantly elevated in the ER-negative and the triple-negative subgroups. These three circRNAs also displayed a desirable potential as diagnostic biomarkers.

Conclusion: This is the first paper that reports the upregulation of circCSPP1 and circSMAD2 in BC and upregulation of circCSPP1 and circNRIP1 in the triple-negative subgroup. Our findings suggest that circCSPP1, circNRIP1, and circSMAD2 may serve as promising diagnostic biomarkers for BC. Identifying the downstream pathways regulated by these circRNAs could lead to the discovery of new therapeutic targets.
Keywords
Breast neoplasms
Circular RNA
Triple Negative breast neoplasms
Tumor biomarkers