Use of ovary culture techniques in reproductive toxicology

Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved. Acknowledgements The author's studies in this field are supported by MRC grants G1002118 (NS and RAA) and G110357 (RAA), MR/L010011/1 (PAF), the European Community's Seventh Framework Programme (FP7/2007–2013) under grant agreement no. 212885 (PAF) and the Wellcome Trust (080388 to PAF). AS was funded by a BBSRC CASE Studentship co-funded by AstraZeneca.Peer reviewedPublisher PD

Hydroxylase Activity of ASPH Promotes Hepatocellular Carcinoma Metastasis Through Epithelial-to-Mesenchymal Transition Pathway

Over-expression of aspartyl (asparagynal)-β-hydroxylase (ASPH) contributes to hepatocellular carcinoma (HCC) invasiveness, but the role of ASPH hydroxylase activity in this process remains to be defined. As such, the current study investigated the role of ASPH hydroxylase activity in downstream signalling of HCC tumorgenesis and, specifically, metastasis development. Over-expression of wild-type ASPH, but not a hydroxylase mutant, promoted HCC cell migration in vitro, as well as intrahepatic and distant metastases in vivo. The enhanced migration and epithelial to mesenchymal transition (EMT) activation was notably absent in response to hydroxylase activity blockade. Vimentin, a regulator of EMT, interacted with ASPH and likely mediated the effect of ASPH hydroxylase activity with cell migration. The enhanced hydroxylase activity in tumor tissues predicted worse prognoses of HCC patients. Collectively, the hydroxylase activity of ASPH affected HCC metastasis through interacting with vimentin and regulating EMT. As such, ASPH might be a promising therapeutic target of HCC. Keywords: ASPH, Hydroxylase, Hepatocellular carcinoma, Metastasis, Epithelial-mesenchymal transition, Vimenti

Correlation between intratumoral AXL expression and clinicopathologic features.

<p><b>Abbreviations</b>: AFP, alpha-fetoprotein; HBsAg, hepatitis B surface antigen; HBeAg, hepatitis E antigen; PLT, platelet count; TBIL total bilirubin; ALB, albumin; ALT, alanine aminotransferase; PVTT, portal vein tumor thrumbus; MVI, microvascular invasion.</p

AXL expression and its association with prognosis in HCC patients.

<p>A, B: Kaplan-Meier curve of TTR and overall survival rates in patients with positive and negative AXL staining, respectively.</p

Effects of AXL expression on cell growth and invasiveness of SMMC-7721.

<p>A: Western blot analysis of cell lysate expressing the control vector (mock), the wild-type AXL (AXL-WT, stable transfection), and RNAi AXL (Si-AXL, transient transfection after 72 hours). B: Cell growth curve of cells transfected with constructs as indicated by MTT assay. C, D: Representative plots of flow cytometry and statistical results of cell apoptosis of cells transfected by constructs as indicated by propidium iodide and annexin V-FITC staining. Bars, ±SD. *, <i>P</i><0.05 versus mock, **, <i>P</i><0.001 versus mock. E: Statistical results of cells transfected with constructs as indicated in transwell assay. *, <i>P</i><0.05 versus mock, **, <i>P</i><0.001 versus mock. F: Representative image of cells transfected with constructs as indicated in the wound healing assay. Mock: control vector; AXL-WT: the wild-type AXL; si-Axl: transient transfection after 72 hours.</p