title: ppGalNAc T1 as a potential novel marker for human bladder cancer.
abstract: OBJECTIVE
  To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo.
  
  
  METHODS
  DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes.
  
  
  RESULTS
  ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%.
  
  
  CONCLUSIONS
  ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.
authors: Department of Urology, The Second Affiliated Hospital of Kunming Medical University, Yunnan Institute of Urology, Kunming, China E-mail : jiansongwang@126.com.
authors: Ming-Xia
authors: MX
authors: Ding
authors: Hai-Feng
authors: HF
authors: Wang
authors: Jian-Song
authors: JS
authors: Wang
authors: Hui
authors: H
authors: Zhan
authors: Yi-Gang
authors: YG
authors: Zuo
authors: De-Lin
authors: DL
authors: Yang
authors: Jing-Yu
authors: JY
authors: Liu
authors: Wei
authors: W
authors: Wang
authors: Chang-Xing
authors: CX
authors: Ke
authors: Ru-Ping
authors: RP
authors: Yan
mesh_terms: D000818:Animals; D017209:Apoptosis; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D002465:Cell Movement; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D017350:N-Acetylgalactosaminyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D001743:Urinary Bladder / Q000378:metabolism*; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D014408:Biomarkers, Tumor; D017350:N-Acetylgalactosaminyltransferases; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000097763:Polypeptide N-acetylgalactosaminyltransferase