MiR-16-5p is frequently down-regulated in astrocytic gliomas and modulates glioma cell proliferation, apoptosis and response to cytotoxic therapy.

AIMS: Aberrant expression of microRNAs (miRNAs) is frequent in various cancers including gliomas. We aimed to characterize the role of miR-16-5p as a candidate tumour suppressor miRNA in gliomas. METHODS: Real-time PCR-based approaches were used for miRNA and mRNA expression profiling of glioma and non-neoplastic brain tissues as well as glioma cell lines. Protein levels were determined by Western blotting. In vitro analyses were performed following overexpression of miR-16-5p, trichostatin A (TSA) treatment, and siRNA-mediated knock-down of HDAC3 in glioma cells. Effects of miR-16-5p on glioma cell viability, apoptosis and response to irradiation and temozolomide (TMZ) were assessed. RESULTS: Expression of miR-16-5p was reduced relative to control brain tissue in isocitrate dehydrogenase (IDH)-mutant astrocytomas of World Health Organization (WHO) grades II, III and IV, and a subset of IDH-wildtype glioblastomas WHO grade IV. MiR-16-5p expression was lower in IDH-mutant than in IDH-wildtype gliomas, and down-regulated in IDH-wildtype glioma lines. MiR-16-5p overexpression reduced expression of important cell cycle and apoptosis regulators in glioma cells, including CDK6, CDC25A, CCND3, CCNE1, WEE1, CHEK1, BCL2 and MCL1. In line, CDK6, WEE1, CHEK1, BCL2 and MCL1 transcript levels were increased in WHO grade III or IV gliomas. TSA treatment and HDAC3 knockdown in glioma cells induced miR-16-5p up-regulation and reduced expression of its targets. Moreover, miR-16-5p overexpression inhibited proliferation and induced apoptosis in various glioma cell lines and increased sensitivity of A172 glioma cells to irradiation and TMZ. CONCLUSION: Reduced expression of miR-16-5p contributes to glioma cell proliferation, survival and resistance to cytotoxic therapy.

Interobserver agreement and accuracy of non-invasive diagnosis of endometriosis by transvaginal sonography.

OBJECTIVES: To evaluate interobserver agreement and accuracy of transvaginal sonography (TVS) in diagnosing deep infiltrating endometriosis (DIE) and endometriomas. METHODS: A total of 67 consecutive patients referred to a pelvic pain clinic and scheduled for laparoscopy were enrolled in the study between January 2013 and January 2014. Patients were independently examined prospectively by two experienced sonographers (Observers A and B) who were blinded to the other's results. For the two observers, Gwet's first-order agreement coefficient (Gwet's AC1) was used to calculate interobserver agreement and diagnostic accuracy, as well as sensitivity, specificity, positive (PPV) and negative (NPV) predictive values using TVS, as compared to laparoscopy, for diagnosing DIE and endometriomas. RESULTS: Among the 67 patients enrolled, 65 were analyzed. For the diagnosis of DIE and endometriomas by TVS, the level of agreement (Gwet's AC1) between Observers A and B and sensitivity/specificity values for the respective Observers were, by site: vagina (Gwet's AC1, 0.933; 62%/94% and 82%/94%), bladder (Gwet's AC1, 1.00; 67%/97% and 67%/97%), uterosacral ligaments (Gwet's AC1, 0.84; 73%/83% and 53%/90%), adnexa (Gwet's AC1, 0.95; 71%/93% and 71%/93%), rectovaginal septum (Gwet's AC1, 0.95; 40%/90% and 33%/87%) and rectosigmoid (Gwet's AC1, 0.98; 93%/96% and 94%/98%) which reflected high interobserver agreement. With the exception of sensitivity of diagnosis of DIE affecting the RVS, similar results were observed when TVS was compared with laparoscopy. CONCLUSIONS: TVS is a highly accurate and reproducible method for non-invasive diagnosis of DIE by well-trained professionals.