Increased MicroRNA Levels in Women With Polycystic Ovarian Syndrome but Without Insulin Resistance: A Pilot Prospective Study

Butler, Alexandra E.; Ramachandran, Vimal; Cunningham, Thomas Keith; David, Rachel; Gooderham, Nigel J.; Benurwar, Manasi; Dargham, Soha R.; Hayat, Shahina; Sathyapalan, Thozhukat; Najafi‐Shoushtari, S. Hani; Atkin, Stephen L.

doi:10.3389/fendo.2020.571357

Cited by 16 publications

(11 citation statements)

References 52 publications

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“…Let-7b-3p, previously known as let-7b, was conventionally regarded as the passenger strand was considered to have no biological function. In recent years, however, relevant studies have revealed let-7b-3p to play a significant role in various of diseases (13)(14)(15). Despite this, the specific role and mechanism of let-7b-3p in LUAD remains unclear.…”

Section: Original Articlementioning

confidence: 99%

Let-7b-3p inhibits tumor growth and metastasis by targeting the BRF2-mediated MAPK/ERK pathway in human lung adenocarcinoma

Li¹,

Dong²,

Lü³

et al. 2021

Transl Lung Cancer Res

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Background: Lung cancer is a malignant tumor with the highest morbidity and mortality rates worldwide, of which lung adenocarcinoma (LUAD) is the most common subtype. Overall, current treatments of LUAD are not satisfactory; therefore, novel targets need to be explored. Let-7b-3p is an important member of the let-7 family of microRNAs (miRNAs), and has not been studied separately in LUAD. This study aimed to investigate the role and molecular mechanism of let-7b-3p in LUAD.Methods: Herein, let-7b-3p expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and fluorescence in situ hybridization (FISH) assays. MTT, colony formation assay, flow cytometry analysis, wound-healing, Transwell and in vivo experiments were conducted to assess let-7b-3p's function in LUAD. The downstream target TFIIB-related factor 2 (BRF2) was predicted using bioinformatics analyses and confirmed by dual-luciferase reporter assay and rescue experiments. Additionally, BRF2 was found to affect the MAPK/ERK pathway through transcriptome sequencing analysis and western blot (WB) assay.Results: Let-7b-3p is downregulated in LUAD cells and tissue samples and low let-7b-3p expression is correlated with a poor prognosis in LUAD patients. Let-7b-3p suppresses the proliferation and metastasis of LUAD cells both in vivo and in vitro by directly targeting the BRF2-mediated MAPK/ERK pathway.Conclusions: Let-7b-3p inhibits the development of LUAD and is an ideal novel therapeutic target for the treatment of LUAD.

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Section: Original Articlementioning

confidence: 99%

Let-7b-3p inhibits tumor growth and metastasis by targeting the BRF2-mediated MAPK/ERK pathway in human lung adenocarcinoma

Li¹,

Dong²,

Lü³

et al. 2021

Transl Lung Cancer Res

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“…Down‐regulated in young women with PCOS ( n = 24). 43 No difference between obese ( n = 21) and NW ( n = 19) women but correlated with waist circumference 102 …”

Section: Resultsmentioning

confidence: 99%

“…Up‐regulated in PCOS 43 and relapsing multiple sclerosis (MS), may be involved in inflammatory pathways 44 …”

Section: Resultsmentioning

confidence: 99%

MicroRNAs in obesity, sarcopenia, and commonalities for sarcopenic obesity: a systematic review

Dowling

Duseja

Vilaça

et al. 2022

J cachexia sarcopenia muscle

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Sarcopenic obesity is a distinct condition of sarcopenia in the context of obesity, with the cumulative health risks of both phenotypes. Differential expression of microRNAs (miRNAs) has been reported separately in people with obesity and sarcopenia and may play a role in the pathogenesis of sarcopenic obesity. However, this has not been explored to date. This study aimed to identify differentially expressed miRNAs reported in serum, plasma, and skeletal muscle of people with obesity and sarcopenia and whether there are any commonalities between these conditions. We performed a systematic review on Embase and MEDLINE (PROSPERO, CRD42020224486) for differentially expressed miRNAs (fold change >1.5 or P-value <0.05) in (i) sarcopenia or frailty and (ii) obesity or metabolic syndrome. The functions and targets of miRNAs commonly changed in both conditions, in the same direction, were searched using PubMed. Following deduplication, 247 obesity and 42 sarcopenia studies were identified for full-text screening. Screening identified 36 obesity and 6 sarcopenia studies for final inclusion. A total of 351 miRNAs were identified in obesity and 157 in sarcopenia. Fifty-five miRNAs were identified in both obesity and sarcopenia-by sample type, 48 were found in plasma and one each in serum and skeletal muscle. Twenty-four miRNAs were identified from 10 of the included studies as commonly changed in the same direction (22 in plasma and one each in serum and skeletal muscle) in obesity and sarcopenia. The majority of miRNA-validated targets identified in the literature search were members of the phosphoinositide 3-kinase/protein kinase B and transforming growth factor-β signalling pathways. The most common targets identified were insulin-like growth factor 1 (miR-424-5p, miR-483-3p, and miR-18b-5p) and members of the SMAD family (miR-483-3p, miR-92a-3p, and miR-424-5p). The majority of commonly changed miRNAs were involved in protein homeostasis, mitochondrial dynamics, determination of muscle fibre type, insulin resistance, and adipogenesis. Twenty-four miRNAs were identified as commonly dysregulated in obesity and sarcopenia with functions and targets implicated in the pathogenesis of sarcopenic obesity. Given the adverse health outcomes associated with sarcopenic obesity, understanding the pathogenesis underlying this phenotype has the potential to lead to effective screening, monitoring, or treatment strategies. Further research is now required to confirm whether these miRNAs are differentially expressed in older adults with sarcopenic obesity.

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“…Due to the low concentration of cell-free circulating RNAs in biofluids such as plasma, precise estimation of RNA concentration is challenging. In this study, RT was performed using equal volume of RNA originating from the equal volume of plasma as described by previous studies (Blondal et al, 2013;Cook et al, 2019;Butler et al, 2020). RT for plasma miRNAs was performed using 2 µl RNA template in 10 µl total RT reaction volume using miRCURY LNA ™ Universal RT miRNA cDNA synthesis kit II (Exiqon), with the addition of 0.625 µl spike-in of synthetic miRNA, UniSp6 (10 8 copies/µl) (Exiqon) for normalization of variation in the RT process.…”

Section: Rna Extraction and Rt-qpcrmentioning

confidence: 99%

Whole Blood Holding Time Prior to Plasma Processing Alters microRNA Expression Profile

et al. 2022

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MicroRNAs (miRNAs) can exhibit aberrant expression under different physiological and pathological conditions. Therefore, differentially expressed circulating miRNAs have been a focus of biomarker discovery research. However, the use of circulating miRNAs comes with challenges which may hinder the reliability for their clinical application. These include varied sample collection protocols, storage times/conditions, sample processing and analysis methods. This study focused on examining the effect of whole blood holding time on the stability of plasma miRNA expression profiles. Whole blood samples were collected from healthy pregnant women and were held at 4°C for 30 min, 2 h, 6 h or 24 h prior to processing for plasma isolation. Plasma RNA was extracted and the expression of 179 miRNAs were analyzed. Unsupervised principal component analysis demonstrated that whole blood holding time was a major source of variation in miRNA expression profiles with 53 of 179 miRNAs showing significant changes in expression. Levels of specific miRNAs previously reported to be associated with pregnancy-associated complications such as hsa-miR-150-5p, hsa-miR-191-5p, and hsa-miR-29a-3p, as well as commonly used endogenous miRNA controls, hsa-miR-16-5p, hsa-miR-25-3p, and hsa-miR-223-3p were significantly altered with increase in blood holding time. Current protocols for plasma-based miRNA profiling for diagnostics describe major differences in whole blood holding periods ranging from immediately after collection to 26 h after. Our results demonstrate holding time can have dramatic effects on analytical reliability and reproducibility. This highlights the importance of standardization of blood holding time prior to processing for plasma in order to minimize introduction of non-biological variance in miRNA profiles.

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Increased MicroRNA Levels in Women With Polycystic Ovarian Syndrome but Without Insulin Resistance: A Pilot Prospective Study

Cited by 16 publications

References 52 publications

Let-7b-3p inhibits tumor growth and metastasis by targeting the BRF2-mediated MAPK/ERK pathway in human lung adenocarcinoma

Let-7b-3p inhibits tumor growth and metastasis by targeting the BRF2-mediated MAPK/ERK pathway in human lung adenocarcinoma

MicroRNAs in obesity, sarcopenia, and commonalities for sarcopenic obesity: a systematic review

Whole Blood Holding Time Prior to Plasma Processing Alters microRNA Expression Profile

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