Interleukin-6-driven progranulin expression increases cholangiocarcinoma growth by an Akt-dependent mechanism

Abstract

Background and objectives Cholangiocarcinoma is a devastating cancer of biliary origin with limited treatment options. The growth factor, progranulin, is overexpressed in a number of tumours. The study aims were to assess the expression of progranulin in cholangiocarcinoma and to determine its effects on tumour growth. Methods The expression and secretion of progranulin were evaluated in multiple cholangiocarcinoma cell lines and in clinical samples from patients with cholangiocarcinoma. The role of interleukin 6 (IL-6)-mediated signalling in the expression of progranulin was assessed using a combination of specific inhibitors and shRNA knockdown techniques. The effect of progranulin on proliferation and Akt activation and subsequent effects of FOXO1 phosphorylation were assessed in vitro. Progranulin knockdown cell lines were established, and the effects on cholangiocarcinoma growth were determined. Results Progranulin expression and secretion were upregulated in cholangiocarcinoma cell lines and tissue, which were in part via IL-6-mediated activation of the ERK1/2/RSK1/C/EBPβ pathway. Blocking any of these signalling molecules, by either pharmacological inhibitors or shRNA, prevented the IL-6-dependent activation of progranulin expression. Treatment of cholangiocarcinoma cells with recombinant progranulin increased cell proliferation in vitro by a mechanism involving Akt phosphorylation leading to phosphorylation and nuclear extrusion of FOXO1. Knockdown of progranulin expression in cholangiocarcinoma cells decreased the expression of proliferating cellular nuclear antigen, a marker of proliferative capacity, and slowed tumour growth in vivo. Conclusions Evidence is presented for a role for progranulin as a novel growth factor regulating cholangiocarcinoma growth. Specific targeting of progranulin may represent an alternative for the development of therapeutic strategies.

Bile Acid Signaling Is Involved in the Neurological Decline in a Murine Model of Acute Liver Failure

“…RNA was extracted from tissues or cells, and real-time PCR was performed, as previously described, 30 using commercially available primers designed against mouse ASBT, FXR, SHP, and glyceraldehyde-3-phosphate dehydrogenase (SABiosciences). A DDCT analysis was performed using vehicle-treated tissue or untreated primary neurons as controls for subsequent experiments.…”

“…FXR nuclear translocation was assessed by immunofluorescence microscopy, as described previously, 30 and neurons were counterstained with the nuclear stain DAPI. In parallel, SHP expression was assessed by real-time PCR, as described previously.…”

Human liver flukes

“…Extracellular vesicles containing excretorysecretory products, are taken up by cholangiocytes, and induce release of IL-6, as well as changes in protein expression associated with cancer (51) . IL-6 drives release of human pro-granulin, which is associated with development of many cancers, including cholangiocarcinoma (52) , and may represent a biomarker for development of advanced periportal fibrosis and cholangiocarcinoma (36,50) .…”

Role of transglutaminase 2 in PAC1 receptor mediated protection against hypoxia-induced cell death and neurite outgrowth in differentiating N2a neuroblastoma cells

“…Rp-cAMPs (10-100 µM; Werner et al, 2014;Mangiavacchi and Wolf, 2004;Wang et al, 2012;Pratt et al, 2016;Vallejo and Vallejo, 2002); Ro 31-8220 (1-10 µM; Lee et al, 2013;Montejo-López et al, 2016), PD 98059 (10-50 µM; Sutter et al, 2004;Kim et al, 2008), SP 600125 (20 µM; Hah et al, 2013;Kim et al, 2010) and Akt inhibitor XI (1 µM; Frampton et al,2012;Rybchyn et al, 2011 Inhibitor Cocktail 2). Cell lysates were scraped and clarified by centrifugation at 4°C for 10 min at 14000 x g prior to being assayed for TG activity using the biotin-labeled cadaverine incorporation assay (see below).…”