The Diagnostic Efficacy of Plasma miR-362-3p Combined with Doppler Ultrasound in the Diagnosis of Breast Cancer

Summary

The diagnosis rate of breast cancer is relatively low. The fourfold table method was used to analyze the diagnostic value of miR-362-3p combined with Doppler ultrasound for breast cancer. The combination of Doppler ultrasound and miR-362-3p effectively enhanced the diagnostic efficacy, providing a promising method for breast cancer diagnosis.

Abstract

Breast cancer is the second most common malignant tumor in the world, seriously threatening the lives of women and imposing a huge economic burden. This study aims to explore the value of miR-362-3p combined with Doppler ultrasound in the diagnosis of breast cancer. A total of 182 breast cancer patients and 147 patients with benign breast disease were included in this study. The plasma miR-362-3p levels of the patients were measured using Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The chi-square test was used to analyze the differences in the parameters of Doppler ultrasound between the two groups of patients. The receiver operator characteristic (ROC) curve was used to evaluate the diagnostic effectiveness of miR-362-3p, Resistance Index (RI), and Pulsatility Index (PI). The diagnostic efficiency of Doppler ultrasound combined with plasma miR-362-3p in breast cancer was analyzed through the fourfold table method. The miR-362-3p levels were markedly decreased in breast cancer patients. There were significant differences in two-dimensional ultrasound parameters. miR-362-3p, RI, and PI have diagnostic value for breast cancer. The combined diagnostic value of the three factors was higher. Ultrasound combined with miR-362-3p significantly improved the sensitivity, accuracy, and negative predictive value in breast cancer diagnosis. Doppler ultrasound combined with miR-362-3p could enhance the diagnostic efficiency of breast cancer, which may provide a non-invasive and potential auxiliary diagnostic tool for breast cancer.

Introduction

According to the statistics of the International Agency for Research on Cancer, the incidence rate of breast cancer exceeded that of lung cancer in 2020¹. Breast cancer is a malignant tumor that commonly occurs and is a leading cause of cancer-related death in women globally¹. Timely diagnosis of breast cancer is beneficial to the prognosis of patients. Tissue biopsy is the gold standard for diagnosing breast cancer. However, this is a type of invasive diagnosis, and it does not apply to patients with early-stage breast cancer².

Doppler ultrasound is a commonly used ima....

Protocol

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of Affiliated Hospital of Shandong Second Medical University, School of Clinical Medicine, Shandong Second Medical University (approval number: wyfy-2023-ky-124). Informed consent was obtained from all subjects involved in the study. The reagents and the equipment used are listed in the Table of Materials.

1. Experimental subjects

Sample size calculation was performed using G*Power 3.1 software based on an independent sample t-test. A medium eff....

Results

The level of miR-362-3p was significantly decreased in breast cancer patients

The clinical baseline information and clinical pathological characteristics of 182 breast cancer patients and 147 patients with benign breast lesions were compared. The detailed data were summarized in Table 1. The RT-qPCR was used to measure the levels of plasma miR-362-3p in two groups of patients. The results showed that the circulating miR-362-3p expression in breast cancer patie.......

Discussion

Breast cancer is one of the most common types of malignant tumors, becoming a major public health issue threatening human health^19,20. The high incidence and mortality rates not only cause severe psychological trauma to the families of the patients but also bring about a huge burden on society and the economy. Most breast cancer patients have already experienced local invasion or distant metastasis at the time of diagnosis^21,

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
BD Vacutainer K2EDTA Blood Collection Tube (purple cap)	USA	367861	For venous blood collection and plasma preparation for miRNA analysis.
CFX96 Touch Real-Time PCR Detection System	Bio-Rad Laboratories	1855195	96-well 6-channel fluorescence quantitative PCR instrument, using optical fiber conduction and cold CCD imaging, with synchronous fluorescence collection in each well without delay.The dynamic range reaches 10 orders of magnitude, and it is compatible with both conventional qPCR and high-resolution melting curve (HRM) analysis.The software supports RDML format export, is compatible with both research and clinical testing, and is commonly used in experiments such as gene expression quantification, SNP typing, and methylation analysis.
Kirgen 0.1 mL PCR Tubes (or 8-Strip PCR Tubes), Sterile	Shanghai, China	KG0811	Used for preparing and performing qPCR reactions.
Kirgen 1.5 mL Microcentrifuge Tubes, Sterile	Kirgen Biotech Co., Ltd.	KG2911	Used for sample storage, reagent preparation, and centrifugation during miRNA extraction and reverse transcription.
miRcute miRNA Isolation Kit (Spin Column)	Beijing, China	DP501	For the isolation and purification of total RNA including miRNA from human plasma samples, using spin column-based technology.
miRcute Plus miRNA First-Strand cDNA Kit	Beijing, China	KR211	For the reverse transcription of purified miRNA into first-strand cDNA via the poly(A) tailing method.
miRcute Plus miRNA qPCR Kit (SYBR Green)	Beijing, China	FP411	For quantitative real-time PCR detection of miRNA using SYBR Green I-based chemistry.
Roche LightCycler 480 Multiwell Plate 96, White	Mannheim, Germany	4729692001	Used for quantitative real-time PCR assays on the Roche LightCycler 480 instrument.
Siemens Acuson 14L5 / 14L5 SP Linear Array Probe	Siemens Healthineers	10041226	High-frequency linear array probe, with a frequency range of 5–14 MHz, specially designed for the examination of superficial organs such as the breast and thyroid. It features high resolution and excellent tissue contrast, and supports elastography and automatic breast volume scanning.
Siemens Acuson S2000/S3000/S1000 Ultrasound System	Siemens Healthineers	/	The color Doppler ultrasound diagnostic system supports real-time 3D/4D imaging and elasticity imaging functions, and is widely used in multiple departments such as abdomen, cardiovascular, gynecology and obstetrics, and breast.
Veriti 96-Well Thermal Cycler	Thermo Fisher Scientific	4375786	96-channel gradient PCR instrument, Supports gradient temperature optimization; has high temperature control accuracy; used for cDNA synthesis and conventional PCR amplification.