MicroRNAs (miRNAs) are small RNA molecules, with their role in gene silencing and translational repression by binding to target mRNAs. Since it was discovered in 1993, miRNA are found in all eukaryotic cells conserved across the species. In recent years, regulation of miRNAs are extensively studied for their role in biological processes as well as in development and progression of various human diseases including retinal disorder, neurodegenerative diseases, cardiovascular disease and cancer. This chapter summarises miRNA biogenesis and explores their potential roles in a variety of diseases. miRNAs holds huge potential for diagnostic and prognostic biomarkers, and as predictors of drug response.

MiRNA Biogenesis and Regulation of Diseases: An Overview.

microRNAs are promising candidate breast cancer biomarkers due to their cancer-specific expression profiles. However, efforts to develop circulating breast cancer biomarkers are challenged by the heterogeneity of microRNAs in the blood. To overcome this challenge, we aimed to develop a molecular profile of microRNAs specifically secreted from breast cancer cells. Our first step towards this direction relates to capturing and analyzing the contents of exosomes, which are small secretory vesicles that selectively encapsulate microRNAs indicative of their cell of origin. To our knowledge, circulating exosome microRNAs have not been well-evaluated as biomarkers for breast cancer diagnosis or monitoring. Exosomes were collected from the conditioned media of human breast cancer cell lines, mouse plasma of patient-derived orthotopic xenograft models (PDX), and human plasma samples. Exosomes were verified by electron microscopy, nanoparticle tracking analysis, and western blot. Cellular and exosome microRNAs from breast cancer cell lines were profiled by next-generation small RNA sequencing. Plasma exosome microRNA expression was analyzed by qRT-PCR analysis. Small RNA sequencing and qRT-PCR analysis showed that several microRNAs are selectively encapsulated or highly enriched in breast cancer exosomes. Importantly, the selectively enriched exosome microRNA, human miR-1246, was detected at significantly higher levels in exosomes isolated from PDX mouse plasma, indicating that tumor exosome microRNAs are released into the circulation and can serve as plasma biomarkers for breast cancer. This observation was extended to human plasma samples where miR-1246 and miR-21 were detected at significantly higher levels in the plasma exosomes of 16 patients with breast cancer as compared to the plasma exosomes of healthy control subjects. Receiver operating characteristic curve analysis indicated that the combination of plasma exosome miR-1246 and miR-21 is a better indicator of breast cancer than their individual levels. Our results demonstrate that certain microRNA species, such as miR-21 and miR-1246, are selectively enriched in human breast cancer exosomes and significantly elevated in the plasma of patients with breast cancer. These findings indicate a potential new strategy to selectively analyze plasma breast cancer microRNAs indicative of the presence of breast cancer.

/pdf/plasma-exosome-micrornas-are-indicative-of-breast-cancer-3uxo26nipk.pdf

Plasma exosome microRNAs are indicative of breast cancer

Regulation of multidrug resistance by microRNAs in anti-cancer therapy.

https://www.fertstert.org/article/S0015-0282(16)61105-6/pdf

Serum microRNAs as diagnostic markers of endometriosis: a comprehensive array-based analysis

Breast cancer is the second cause of cancer mortality in women globally. Early detection, treatment, and metastasis monitoring are of great importance to favorable prognosis. Although conventional diagnostic methods, such as breast X-ray mammography and image positioning biopsy, are accurate, they could cause radioactive or invasive damage to patients. Liquid biopsy as a noninvasive method is convenient for repeated sampling in clinical cancer prognostic, metastatic evaluation, and relapse monitoring. MicroRNAs encased in exosomes circulating in biofluids are promising candidate cancer biomarkers because of their cancer-specific expression profiles. Here, we report an in situ detection of microRNA-1246 (miR-1246) in human plasma exosomes as breast cancer biomarker by a nucleic acid functionalized Au nanoflare probe. Needing neither time-consuming and costly isolation of exosomes from the plasma sample nor transfection means, the Au nanoflare probe can directly enter the plasma exosomes to generate fluorescent signal quantitatively by specifically targeting miR-1246. Only 40 μL of plasma is needed to incubate 4 h with the probe, giving signal sensitive enough to distinguish samples of breast cancer to normal control. Using plasma miR-1246 level detected by our assay as a marker, we differentiated 46 breast cancer patients from 28 healthy controls with 100% sensitivity and 92.9% specificity at the best cutoff. This simple, accurate, sensitive, and cost-effective liquid biopsy by the Au nanoflare probe is potent to be developed as a noninvasive breast cancer diagnostic assay for clinical adaption.

In Situ Detection of Plasma Exosomal MicroRNA-1246 for Breast Cancer Diagnostics by a Au Nanoflare Probe

A bunch of microRNAs (miRNAs) have been demonstrated to be aberrantly expressed in cancer tumor tissue and serum. The miRNA signatures identified from the serum samples could serve as potential noninvasive diagnostic markers for breast cancer. The role of the miRNAs in cancerigenesis is unclear. In this study, we generated the expression profiles of miRNAs from the paired breast cancer tumors, normal, tissue, and serum samples from eight patients using small RNA-sequencing. Serum samples from eight healthy individuals were used as normal controls. We identified total 174 significantly differentially expressed miRNAs between tumors and the normal tissues, and 109 miRNAs between serum from patients and serum from healthy individuals. There are only 10 common miRNAs. This suggests that only a small portion of tumor miRNAs are released into serum selectively. Interestingly, the expression change pattern of 28 miRNAs is opposite between breast cancer tumors and serum. Functional analysis shows that the differentially expressed miRNAs and their target genes form a complex interaction network affecting many biological processes and involving in many types of cancer such as prostate cancer, basal cell carcinoma, acute myeloid leukemia, and more.

Different miRNA expression profiles between human breast cancer tumors and serum.

The purpose of the present study was to investigate the serum levels of microRNA (miRNA/miR)-382-3p, -598-3p, -1246 and -184 in breast cancer patients and to assess their feasibility as biomarkers for breast cancer screening. Serum samples were obtained from 100 breast cancer patients and 40 age-matched healthy control subjects in Taizhou Central Hospital (Taizhou, Zhejiang, China) between January 2013 and September 2014. The serum expression levels of miR-382-3p, -598-3p, -1246 and -184 were determined by stem-loop reverse transcription-quantitative polymerase chain reaction. Receiver operating characteristic curves were drawn to evaluate the sensitivity and specificity of the serum miRNA expression levels for the screening of breast cancer. miR-382-3p and -1246 were significantly upregulated in the serum of the breast cancer patients, while miR-598-3p and -184 were significantly downregulated. The sensitivity and specificity to detect breast cancer were as follows: miR-382-3p, 52.0 and 92.5%; miR-598-3p, 95.0 and 85.0%; miR-1246, 93.0 and 75.0%; and miR-184, 87.5 and 71.0%, respectively. The expression levels of the four serum miRNAs were not correlated with the patients' clinical stage. In summary, miR-382-3p, -598-3p, -1246 and -184 are all involved in the development of breast cancer, and are promising biomarkers for breast cancer detection.

/pdf/serum-expression-levels-of-microrna-382-3p-598-3p-1246-and-3y3kkuqzfk.pdf

Serum expression levels of microRNA-382-3p, -598-3p, -1246 and -184 in breast cancer patients.

Objective
To explore the effect of extracts of Prunella vulgaris L. on proteome of human lung adenocarcinoma cell line A549 by two-dimensional electrophoresis and mass spectrometry and elucidate the mechanism of anti-lung adenocarcinom effect of Prunella vulgaris L. at the level of proteome.


Methods
The proliferative activity of human lung adenocarcinoma cell line A549 was evaluated by methyl thiazolyl tetrazolium (MTT) colorimetric assay. According to the difference of culture medium, all subjects were divided into the experimental group with culture medium of extracts of Prunella vulgaris L. (300 μg/ml) and the control group with culture medium of DMSO (0.3%). Proteins were isolated by two-dimensional electrophoresis and proteomic maps acquired by silver staining. And proteomic analysis was processed by Image Master 2D Quant Platinum 6.0. The proteins with > 2-fold differences were used to analyze by mass spectrometry and confirmed by Western blot.


Results
The expressions of inositol 1, 4, 5-triphosphate receptor-interacting protein-like 2 precursor, heat shock cognate protein 70, serine-threonine kinase receptor-associated protein, tropomyosin 2(β) isoform 1, cyclin B3, MED12L protein and macrophin 1 isoform 2 were higher in experimental group than those in control group(ratio (medicial/normal) 2.051 93, 1 000 001, 2.203 08, 5.042 01, 15.178 00, 1 000 001, 1 000 001). And the expressions of enolase 1, M2-type pyruvate kinase, heat shock protein 27, Rho GDP-dissociation inhibitor 1, heat shock protein β1, TapasinERP57 heterodimer chain A, inorganic pyrophosphatase and mitochondrial Cysteinyl-tRNA synthetase 2 (putative) were lower in experimental group than those in control group(ratio (medicial/normal) 0.485 18, 0.491 53, 0.465 43, 0.454 71, 0.499 34, 0.450 36, 0.494 62, 0.437 33).


Conclusions
The extracts of Prunella vulgaris L. have multi-target and multi-pathway effects on anti-lung adenocarcinoma. And its possible mechanisms may be due to the regulation of steady state of calcium ion, cell cycle and its steady state and the inhibition of tumor cell proliferation and metastasis.


Key words: 
Prunella; Lung neoplasms; Proteome

Effects of extracts of Prunella Vulgaris L. on proteome of human lung adenocarcinoma cell line A549

The invention discloses a blood serum miRNA (Micro Ribonucleic Acid) marker related to a breast cancer and an application of the marker. The marker is a combination of miR-598-3p or miR-598-3p and CA153. The marker and a primer of the marker can be used for preparing a diagnostic kit for early diagnosis of the human breast cancer.

Jie Zhu

Papers

Different miRNA expression profiles between human breast cancer tumors and serum.

Serum expression levels of microRNA-382-3p, -598-3p, -1246 and -184 in breast cancer patients.

Effects of extracts of Prunella Vulgaris L. on proteome of human lung adenocarcinoma cell line A549

Blood serum marker related to human breast cancer and application of blood serum marker