Hormone replacement therapy enhances IGF-1 signaling in skeletal muscle by diminishing miR-182 and miR-223 expressions : a study on postmenopausal monozygotic twin pairs

MiRNAs are fine-tuning modifiers of skeletal muscle regulation, but knowledge of their hormonal control is lacking. We used a co-twin case-control study design, that is, monozygotic postmenopausal twin pairs discordant for estrogen-based hormone replacement therapy (HRT) to explore estrogen-dependent skeletal muscle regulation via miRNAs. MiRNA profiles were determined from vastus lateralis muscle of nine healthy 54-62-years-old monozygotic female twin pairs discordant for HRT (median 7 years). MCF-7 cells, human myoblast cultures and mouse muscle experiments were used to confirm estrogen's causal role on the expression of specific miRNAs, their target mRNAs and proteins and finally the activation of related signaling pathway. Of the 230 miRNAs expressed at detectable levels in muscle samples, qPCR confirmed significantly lower miR-182, miR-223 and miR-142-3p expressions in HRT using than in their nonusing co-twins. Insulin/IGF-1 signaling emerged one common pathway targeted by these miRNAs. IGF-1R and FOXO3A mRNA and protein were more abundantly expressed in muscle samples of HRT users than nonusers. In vitro assays confirmed effective targeting of miR-182 and miR-223 on IGF-1R and FOXO3A mRNA as well as a dose-dependent miR-182 and miR-223 down-regulations concomitantly with up-regulation of FOXO3A and IGF-1R expression. Novel finding is the postmenopausal HRT-reduced miRs-182, miR-223 and miR-142-3p expression in female skeletal muscle. The observed miRNA-mediated enhancement of the target genes' IGF-1R and FOXO3A expression as well as the activation of insulin/IGF-1 pathway signaling via phosphorylation of AKT and mTOR is an important mechanism for positive estrogen impact on skeletal muscle of postmenopausal women.Peer reviewe

Expression of Trop2 in bladder cancer is modulated by miR125b: in vivo and in vitro analyses

Human trophoblastic cell surface antigen 2 (Trop-2) is a 40-kDa transmembrane glycoprotein, first identified as a cell surface marker for human trophoblast cells (1). Elevated expression of Trop-2 has been shown in several types of epithelial cancers and correlated with tumour aggressive and poor prognosis (2-3). The first aim of this study was to evaluate the variation of the Trop-2 expression in normal urothelium and urothelial bladder cancer. The immunohistochemical results showed an increase of Trop-2 levels in bladder cancer tissues with the increase of the severity of the pathology. Recent data identified Trop-2 as a target for miR-125b suggesting a pos sible role of miR-125b in the modulation of Trop-2 protein expression (4). The second aim was to verify if Trop-2 could be a target for miR-125b in bladder cells and to evaluate the possible role of miR-125b in the modulation of Trop-2 protein expression in normal bladder as well as in urothelial bladder cancer. In vitro we showed a contribution of miR-125b in deregulation of Trop-2 protein expression in a bladder cell line and we found that the expression of miR-125b was inversely correlated with the expression of Trop-2 protein on a cohort of bladder cancer tissues. We concluded to investigate in a larger population the use of Trop-2 and/or miR-125b as potential diagnostic markers in urothelial bladder cancer

Dystrophin Is Required for the Normal Function of the Cardio-Protective KATP Channel in Cardiomyocytes

Duchenne and Becker muscular dystrophy patients often develop a cardiomyopathy for which the pathogenesis is still unknown. We have employed the murine animal model of Duchenne muscular dystrophy (mdx), which develops a cardiomyopathy that includes some characteristics of the human disease, to study the molecular basis of this pathology. Here we show that the mdx mouse heart has defects consistent with alteration in compounds that regulate energy homeostasis including a marked decrease in creatine-phosphate (PC). In addition, the mdx heart is more susceptible to anoxia than controls. Since the cardio-protective ATP sensitive potassium channel (KATP) complex and PC have been shown to interact we investigated whether deficits in PC levels correlate with other molecular events including KATP ion channel complex presence, its functionality and interaction with dystrophin. We found that this channel complex is present in the dystrophic cardiac cell membrane but its ability to sense a drop in the intracellular ATP concentration and consequently open is compromised by the absence of dystrophin. We further demonstrate that the creatine kinase muscle isoform (CKm) is displaced from the plasma membrane of the mdx cardiac cells. Considering that CKm is a determinant of KATP channel complex function we hypothesize that dystrophin acts as a scaffolding protein organizing the KATP channel complex and the enzymes necessary for its correct functioning. Therefore, the lack of proper functioning of the cardio-protective KATP system in the mdx cardiomyocytes may be part of the mechanism contributing to development of cardiac disease in dystrophic patients

Circulating inflamma-miRs in aging and age-related diseases.

none4nopubblicazione scientificaOlivieri F.; Rippo M.R.; Procopio A.D.; Fazioli F.Olivieri, Fabiola; Rippo, Maria Rita; Procopio, Antonio Domenico; Fazioli, Francesc

Natural killer activity in spleens and lymph nodes from patients with Hodgkin's disease.

Natural killer (NK) activity against K-562 myeloid cell line was evaluated in 21 spleens and 14 lymph nodes from patients with Hodgkin's disease (HD). NK activity of eight HD-involved (HD+) spleens [556 lytic units (LU)] was found 5-fold higher than that of 13 HD-uninvolved (HD-) spleens (112 LU) (p less than 0.01). Moreover, NK activity of HD+ spleens was significantly different (p less than 0.05) from that of three spleens involved by non-HD lymphoma (100 LU). NK activity of four spleens from nonneoplastic patients (250 LU) was intermediate between those of HD+ and HD- spleens. Lymph node cells were about 10-fold less cytotoxic. NK activity of seven HD+ lymph nodes (43 LU) was 3-fold higher than that of three lymph nodes involved by non-HD lymphoma (8 LU). However, these differences were not statistically significant. Our data are compatible with increased NK activity in HD+ tissues as well as with depressed NK activity in HD- tissues. The observation that NK activity of peripheral blood leukocytes from nine HD patients (116 LU) (p less than 0.05 only at 100:1 effector:target cell ratio) may support the latter interpretation. Partial characterization of effector cells in HD+ and in HD- spleens indicated that, in both instances NK cells were nonadherent and almost equally distributed between the erythrocyte-negative and -positive cell fractions. Finally, NK activity of both HD+ and HD- spleen cells could be further potentiated in vitro by interferon

Expression Levels and Clinical Significance of miR-21-5p, miR-let-7a, and miR-34c-5p in Laryngeal Squamous Cell Carcinoma

Objective. Altered microRNAs (miRNAs) expression has been found in many cancer types, including laryngeal squamous cell carcinoma (LSCC). The aim of this study was to determine the role and clinical value of three LSCC-related miRs, such as miR-21-5p, miR-let-7a, and miR-34c-5p in a homogeneous cohort of patients with primary LSCC treated by primary surgery. Methods. Expression levels of miR-21-5p, miR-let-7a, and miR-34c-5p were detected in 43 pairs of LSCC and adjacent normal tissues by reverse-transcription quantitative PCR. Overall survival and disease-free survival were evaluated using the Kaplan–Meier method, and multivariate analysis was performed using the Cox proportional hazard analysis. Results. miR-21-5p is significantly upregulated, while miR-let-7a is significantly downregulated in LSCC tumor tissues compared with the corresponding adjacent normal tissues. The downregulation of miR-34c-5p expression significantly correlated with a shorter disease-free survival and, in the multivariate analysis, low miR-34c-5p expression was associated with an increased risk of recurrence. Conclusions. miR-21-5p, miR-let-7a, and miR-34c-5p seem to play a critical role in LSCC carcinogenesis and might have a diagnostic and prognostic clinical value. The miR-let-7a levels could have a predictive role for lymph node metastases and miR-34c-5p might be a promising biomarker of patient outcome

Geomorphic effects caused by heavy rainfall in southern Calabria (Italy) on 30 October–1 November 2015

A severe rainfall event occurred in southern Calabria between 29 October and 2 November 2015, causing two deaths and serious damage to transport infrastructure. Widespread slope erosion and thousands of shallow landslides were triggered on the slopes, combined with flooding and transport of debris along the streams. Rains recorded by the regional gauge network and the national radar monitoring system were analysed by means of Kriging techniques. Ground effects were surveyed in the field, and mapped using post-event air photos taken along the coastal sector. Shallow landslides, soil erosion (including sheet, rill, and gully erosion), flooding, lateral erosion, and debris deposition along streambeds, overflow on lateral slopes, and fan deposition at the mouths of the streams were mapped at 1:60,000 scale. Isohyets of the rainfall event – in terms of cumulative maxima over 24 h – are also shown on the Main Map. In Annex A, a list of notices of processes and/or damage, arranged by municipalities, with coordinates, types of ground effect, and synthetic descriptions, is also provided