Novel breast cancer screening: combined expression of miR-21 and MMP-1 in urinary exosomes detects 95% of breast cancer without metastasis

Wataru Ando,Kiyoshi Kikuchi,Hiroaki Yokomori,Takashi Takaki,Isao Okazaki,Takayuki Uematsu,Katsuya Otori,Yutaka Kohgo,Masaya Sogabe,Shigemi Ishikawa

doi:10.1038/s41598-019-50084-5

Abstract

Serum and tissue miR-21 expression in patients with breast cancer (BC) is a useful biomarker for cancer diagnosis, progression, and treatment. Matrix metalloproteinase-1 (MMP-1) is also important in breast cancer carcinogenesis. However, miR-21 and MMP-1/CD63 in urine exosomes in these patients have not been examined. Urine samples were collected from patients with BC and 26 healthy females. Urinary exosomes were isolated and confirmed by western blotting with anti-CD63 antibody and electron microscopy observation. MiR-21 and MMP-1/CD63 expression was examined by quantitative RT-PCR and western blotting, respectively. Patients with very early stage breast cancer were evaluated. MiR-21 expression in the patients was 0.26 [95% CI: 0.20–0.78], which was significant lower than in the 26 controls (1.00 [95% CI: 1.01–3.37], p = 0.0947). MMP-1/CD63 expression in patients was significantly higher than in controls (1.74 [95% CI: 0.86–5.08] vs 0.535 [95% CI: −0.01–2.81], p = 0.0001). Sensitivity and specificity were 0.708 and 0.783 for miR-21 and 0.792 and 0.840 for MMP-1/CD63, respectively. Sensitivity and specificity of combined expression were 95% and 79%, respectively. The sensitivity of MMP-1/CD63 expression in urinary exosomes was better than that of miR-21 expression. Thus, miR-21 and MMP/CD63 may be useful markers for BC screening.

Highlights

The expression of miR-21 in urinary exosomes of patients with breast cancer (BC) has not been reported because they had not confirmed their samples by CD63 antibody nor by the electron microscopy observation[16]
We previously investigated the mechanism of matrix metalloproteinase-1 (MMP-1) in the formation of liver cirrhosis[19,20,21,22,23] and development of hepatocellular carcinoma[24,25,26]
These cases were detected relatively early because most patients visited the Department of Preventive Medicine of Sanno Medical Center for a health check which included BC screening, and suspicious lesions were detected by breast echography and mammography

Summary

Introduction

In Japan, the prevalence of BC has gradually increased, with the highest incidence rate and the third leading cause of mortality among all cancers[2]. Ductal ca scirrhous ca mucinous ca Median (95% CI). Molecular biology studies showed that exosomes secreted from cancer cells can migrate to distant organs to form niches, leading to metastasis[6,7]. Exosomes contain both microRNAs (miRs) and oncogenes[6,7]. This study was performed to determine the expression levels of both miR-21 and MMP-1/CD63 in urinary exosomes and evaluate these biomarkers for screening of early stage BC

Methods

Results

Conclusion