SULF1/SULF2 splice variants differentially regulate pancreatic tumour growth progression

SULF1/SULF2 reactivation during liver damage and tumour growth

“…All PCR products following 40 amplification cycles were analysed in 2% agarose gels (Gill et al 2014).…”

“…Human liver cell lines Hep3B and PRF5 were grown in Dulbecco‫׳‬s modified Eagle‫׳‬s (DMEM) medium with 10% foetal calf serum as described previously (Gill et al 2014). Over-expression of Sulf1 or Sulf2 variants in both these cell lines was achieved by transfection with Biorad transfectin lipid reagent.…”

“…Such transfected cells were used for in vitro and in vivo assays following 2-6 weeks of growth in G418 containing medium. For in vitro proliferation assays, 10,000 cells/well were seeded in multiple 24-well plates and the trypsinised cells counted after 1, 2, 3, 4 and 5 days of culture or only once after 5 days culture in certain assays (Gill et al 2014). In addition to each sample consisting of a minimum of three replicates for each stage, each experiment was repeated three times.…”

“…Changes in Sulf1/Sulf2 expression in normal and pathological tissues could thus indicate their cell signalling activity in such tissues. The role of Sulf1/Sulf2 in HCC growth, however, is not clear due to some conflicting reports of Sulf1/Sulf2 upregulation (Bret et al 2011;Gill et al 2011;Gill et al 2012;Gill et al 2014;Nawroth et al 2007; LemjabbarAlaoui et al ; Rosen and Lemjabbar-Alaoui 2010) as well as the downregulation of such enzymes in HCC (Lai et al 2008a;Lai et al 2006;Xu G et al 2014). This therefore raises the question as to whether Sulf1 or Sulf2 enzymes are preferentially expressed in normal adult or fetal liver or tumour tissues.…”

“…This therefore raises the question as to whether Sulf1 or Sulf2 enzymes are preferentially expressed in normal adult or fetal liver or tumour tissues. This study was undertaken to determine if it is the normal or the tumour tissues that express high levels of Sulf1 or Sulf2 since a number of studies have reported the upregulation of Sulf1 and Sulf2 enzymes in tumours (Kroy DC et al 2010;Gill et al 2012;Gill et al 2014;Bret et al 2011;Nawroth et al 2007) while others have reported the down-regulation of these enzymes in tumour tissues (Lai et al 2006;Lai et al 2008b;Xu G et al 2014) compared with normal tissues thus implying that Sulfs and particularly Sulf1 is a tumour suppressor rather than a tumour growth enhancer. Since HCC develops over a long time period and is often preceded by additional steps such as hepatitis and cirrhosis, this analysis also included some other liver samples likely to activate cell signalling in response to liver damage-induced regeneration.…”