lncRNA-FMR6 directly binds SAV1 to increase apoptosis of granulosa cells in premature ovarian failure.

BACKGROUND: A regulatory mechanism of lncRNA binding to protein has been detected in premature ovarian failure (POF). Therefore, this study was expected to illustrate the mechanism of lncRNA-FMR6 and SAV1 regulating POF. METHODS: Follicular fluid and ovarian granulosa cells (OGCs) from POF patients and healthy volunteers were collected. Using RT-qPCR and western blotting, lncRNA-FMR6 and SAV1 expression were detected. KGN cells were cultured, and the subcellular localization analysis of lncRNA-FMR6 was carried out. In addition, KGN cells were treated with lncRNA-FMR6 knockdown/overexpression or SAV1 knockdown. Then, cell optical density (proliferation), apoptosis rate, Bax and Bcl-2 mRNA expression were explored by CCK-8, caspase-3 activity, flow cytometry and RT-qPCR analysis. By performing RIP and RNA pull-down experiments, the interactions among lncRNA-FMR6 and SAV1 was investigated. RESULTS: Up-regulation of lncRNA-FMR6 was shown in follicular fluid and OGCs of POF patients, and ectopic overexpression of lncRNA-FMR6 promoted KGN cells apoptosis and inhibited proliferation. lncRNA-FMR6 was localized in the cytoplasm of KGN cells. SAV1 bounding to lncRNA-FMR6 was negatively regulated by lncRNA-FMR6, and was down-regulated in POF. SAV1 knockdown promoted KGN cells proliferation and inhibited apoptosis, and partially eliminated the effect of lncRNA-FMR6 low expression on KGN cells. CONCLUSION: Overall, lncRNA-FMR6 accelerates POF progression by binding to SAV1.

The possible involvement of circRNA DMNT1/p53/JAK/STAT in gestational diabetes mellitus and preeclampsia.

Circular RNA (circRNA) plays an important role in biological processes of gestational diabetes mellitus (GDM) and preeclampsia (PE). However, the mechanisms for circRNA DMNT1 (circ-DMNT1) in GDM and PE remain unclarified. The expression levels of circ-DMNT1 and p53 in GDM and PE were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis. When the expression of circ-DMNT1 or p53 was abnormal, cell counting kit-8 (CCK-8) assay, bromodeoxyuridine (BrdU) staining, flow cytometry, cell scratch, and Transwell assays were used to assess cell viability, proliferation, cell cycle, apoptosis, migration, and invasion of trophoblast cells, respectively. Subsequently, the binding relationship between circ-DMNT1 and p53 was verified by RNA pull-down and RIP analysis, followed by the determination of JAK/STAT pathway-related protein expression levels using western blot analysis. Both circ-DMNT1 and p53 were highly expressed in GDM and PE. Upregulation of circ-DMNT1 or p53 inhibited trophoblast cell viability, proliferation, migration, and invasion, meanwhile promoting cell apoptosis but blocking cell cycle progression. However, downregulation of circ-DMNT1 or p53 induced trophoblast cell survival. In GDM and PE, circ-DMNT1 activated the JAK/STAT pathway by binding to p53, which resulted in increased expression levels of p-JAK and p-STAT. The results suggested that circ-DMNT1 was involved in the deterioration of GDM and PE, possibly through inducing p53 expression and activating the JAK/STAT signaling pathway.

A hairpin probe-mediated DNA circuit for the detection of the mecA gene of Staphylococcus aureus based on exonuclease III and DNAzyme-mediated signal amplification.

A simple, highly sensitive biosensor for S. aureus detection is becoming increasingly important in human health and safety. In this work, a hairpin probe-mediated DNA circuit for the detection of the mecA gene of S. aureus was reported cascading Exo III-assisted cycling signal amplification and the DNAzyme-mediated cleavage reaction. In the presence of the target mecA gene, the recognition and hybridization between HP1 and mecA can trigger Exo III and DNAzyme-mediated signal amplification and further release numerous ATMND, resulting in an enhanced fluorescence response, which serves as a response signal for the fluorescence detection of mecA gene. This biosensor enables the sensitive and specific detection of the mecA gene, showing a linear response ranging from 1 fM to 1 nM with a detection limit of 0.5 fM. Moreover, this fluorescence assay has been applied for the analysis of clinical samples with satisfactory recovery. Importantly, this universal platform can be further extended for the analysis of other targets by alternating the corresponding recognition unit, which holds much promise in point-of-care testing for bacterial analysis.

miR-130b-3p is high-expressed in polycystic ovarian syndrome and promotes granulosa cell proliferation by targeting SMAD4.

BACKGROUND: Being one of the most prevalent metabolic and endocrine disorders, Polycystic Ovary Syndrome (PCOS) has been proven to be associated with microRNA-130b-3p (miR-130b-3p). However, the exact role played by miR-130b-3p in the pathogenesis and progression of PCOS remains unknown. Thus, this article is focused on elucidating the function of miR-130b-3p in the pathogenesis of PCOS. METHODS: The expression levels of miR-130b-3p and SMAD4 in tissues and cells responsible for the development of PCOS were determined by RT-qPCR and western blot. A miR-130b-3p mimic/inhibitor or si-SMAD4 were transfected into KGN cells. The cell viability was detected by CCK-8 and EDU methods. The activity of caspase-3 was measured by caspase-3 analysis. Subsequently, apoptosis and the cell cycle were measured via flow cytometry. The correlation between SMAD4 and miR-130b-3p was confirmed using an RNA pull-down assay and a dual luciferase reporter system assay. RESULTS: MiR-130b-3p was upregulated in the KGN cells and ovarian granulosa cells (GCs) of PCOS patients. It was found that miR-130b-3p overexpression or SMAD4 silencing can promote KGN cell proliferation and positive EDU rates, induce S phase arrest, inhibit apoptosis and caspase-3 activity. On the other hand, miR-130b-3p inhibitors reduce KGN cell proliferation, inhibit apoptosis and reverse the effect of si-SMAD4. CONCLUSION: MiR-130b-3p directly interacts with SMAD4 to induce KGN cell proliferation, inhibit apoptosis, suggesting that miR-130b-3p expression is positively correlated with the development of PCOS. This may serve as new evidence for the abnormal proliferation of GCs in PCOS.