title: Identification of metastasis-associated proteins involved in gallbladder carcinoma metastasis by proteomic analysis and functional exploration of chloride intracellular channel 1.
abstract: Advanced gallbladder cancer has an extremely poor prognosis because of metastasis. Identification of metastasis-related biomarkers is essential to improve patient survival. In the present study, metastasis-associated proteins were identified by comparative proteomic analysis and the metastasis-related function of the candidate protein, chloride intracellular channel 1 (CLIC1), was further elucidated. Two cell lines with high or low metastatic potential (termed GBC-SD18H and GBC-SD18L, respectively), originating from the same parental gallbladder carcinoma GBC-SD cell line, were identified by spontaneous metastasis in vivo and characterized by metastatic phenotypes analysis in vitro. Subsequently, a proteomic approach comprised of two-dimensional gel electrophoresis analysis and mass spectroscopy was used to identify and compare the protein expression patterns between GBC-SD18L and GBC-SD18H. Twenty-six proteins were identified and further verified by one-dimensional Western blotting and semiquantitative reverse transcriptase polymerase chain reaction analysis. It was determined that CLIC1, ezrin, vimentin, annexin A3, WD repeat domain 1, triosephosphate isomerase, C1-tetrahydrofolate synthase, Rho GDP-dissociation inhibitor 1, T-complex protein 1, heterogeneous nuclear ribonucleoprotein K, glutamate dehydrogenase 1, proteasome activator complex subunit 3 and Rab GDP-dissociation inhibitor beta were significantly up-regulated in the highly metastatic GBC-SD18H cell line compared to the poorly metastatic GBC-SD18L cell line. However, phosphoglycerate kinase 1 and programmed cell death protein 8 were significantly down-regulated in the highly metastatic GBC-SD18H cell line compared to GBC-SD18L. Considering that CLIC1 was profuse in highly metastatic GBC-SD18H but scarce in poorly metastatic GBC-SD18L, the association of CLIC1 with metastasis was further elucidated by the overexpression and RNA interference of CLIC1 in GBC-SD18L cells and GBC-SD18H cells, respectively. The results demonstrated that the overexpression of CLIC1 promoted cell motility and invasion of GBC-SD18L in vitro, while RNA interference of CLIC1 remarkably decreased cell motility and invasive potency of GBC-SD18H in vitro, indicating that CLIC1 might play an important role in metastasis of gallbladder carcinoma.
authors.affiliation: Department of Surgery, Second Affiliated Hospital, Zhejiang University, Hangzhou, China.
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Wang
authors.forename: Shu-You
authors.initials: SY
authors.lastname: Peng
authors.forename: Jiang-Tao
authors.initials: JT
authors.lastname: Li
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Zhi-Ping
authors.initials: ZP
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Cheng
authors.forename: De-Qing
authors.initials: DQ
authors.lastname: Cheng
authors.forename: Wei-Hong
authors.initials: WH
authors.lastname: Weng
authors.forename: Xiang-Song
authors.initials: XS
authors.lastname: Wu
authors.forename: Xiao-Zhou
authors.initials: XZ
authors.lastname: Fei
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Quan
authors.forename: Ji-Yu
authors.initials: JY
authors.lastname: Li
authors.forename: Song-Gang
authors.initials: SG
authors.lastname: Li
authors.forename: Ying-Bin
authors.initials: YB
authors.lastname: Liu
mesh_terms: D000818:Animals; D002277:Carcinoma / Q000473:pathology / Q000556:secondary*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D005706:Gallbladder Neoplasms / Q000473:pathology*; D020869:Gene Expression Profiling; D006801:Humans; D008113:Liver Neoplasms / Q000556:secondary; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness / Q000503:physiopathology*; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D009368:Neoplasm Transplantation; D040901:Proteomics; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000502:physiology; D013047:Specific Pathogen-Free Organisms
chemical_list: C501966:CLIC1 protein, human; D018118:Chloride Channels; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins