title: Latexin inhibits the proliferation of CD133+ miapaca-2 pancreatic cancer stem-like cells.
abstract: BACKGROUND
  An increasing number of evidence suggests that pancreatic cancer contains cancer stem cells (CSCs), which may be relevant to the resistance of chemotherapy. Latexin (Lxn) is a negative regulator of stem cell proliferation and we investigate the effects of Lxn on CD133+ pancreatic cancer stem-like cells.
  
  
  METHODS
  CD133+ miapaca-2 cells, a human pancreatic carcinoma cell line, were isolated and sorted by magnetic activated cell sorting and flow cytometry. The capacity for self-renewal, proliferation, and tumorigenicity of CD133+ miapaca-2 cells was determined by the floating spheres test and tumor xenograft assays. Protein and mRNA expression of Lxn in CD133+ and CD133- miapaca-2 cells were detected by Western blotting and qRT-PCR, respectively. After CD133+ miapaca-2 cells were treated with Lxn in serum-free medium (SFM), cell proliferation was assayed with a Cell Counting Kit 8 (CCK-8) and apoptosis was analyzed by flow cytometry. The protein and mRNA expression levels of Bcl-2, bax, and c-myc were also analyzed.
  
  
  RESULTS
  We successfully isolated CD133+ miapaca-2 cells that exhibited the capacity for self-renewal in SFM, a proliferation potential in DMEM supplemented with FBS, and high tumorigenicity in nude mice. Lxn protein and mRNA expression levels in CD133+ miapaca-2 cells were significantly lower than those in CD133- cells. Lxn-treated CD133+ miapaca-2 cells exhibited increased apoptosis and low proliferation activity, down-regulation of Bcl-2 and c-myc expression, and up-regulation of Bax expression in a dose-dependent manner.
  
  
  CONCLUSIONS
  Lxn induces apoptosis and inhibits the proliferation of CD133+ miapaca-2 cells. These changes are associated with down-regulation of Bcl-2 and c-myc and up-regulation of Bax.
authors.affiliation: Department of Gastroenterology, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. xuezhanxiong@163.com.
authors.forename: Zhan-Xiong
authors.initials: ZX
authors.lastname: Xue
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. ald433@163.com.
authors.forename: Ji-Hang
authors.initials: JH
authors.lastname: Zheng
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. zhe_zhi2000@aliyun.com.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Zheng
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. cjli2001@tom.com.
authors.forename: Jing-Li
authors.initials: JL
authors.lastname: Cai
authors.affiliation: Departments of Gastroenterology and Hepatology, Jinhua Municipal Central Hospital, Jinhua Hospital of Zhejiang University, Jinhua, 321000, Zhejiang Province, China. 530392865@qq.com.
authors.forename: Xiao-Hua
authors.initials: XH
authors.lastname: Ye
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. wangcheng200617@163.com.
authors.forename: Cheng
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. fame198288@126.com.
authors.forename: Wei-Jian
authors.initials: WJ
authors.lastname: Sun
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. 47487955@qq.com.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. 4574270@qq.com.
authors.forename: Ming-Dong
authors.initials: MD
authors.lastname: Lu
authors.affiliation: Department of General Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. lipihong@163.com.
authors.forename: Pi-Hong
authors.initials: PH
authors.lastname: Li
authors.affiliation: Department of Gastroenterology, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, China. czz77@sina.com.
authors.forename: Zhen-Zhai
authors.initials: ZZ
authors.lastname: Cai
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D000941:Antigens / Q000096:biosynthesis / Q000235:genetics*; D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023041:Xenograft Model Antitumor Assays
chemical_list: D000071916:AC133 Antigen; D000941:Antigens; D015703:Antigens, CD; D006023:Glycoproteins; C086812:Lxn protein, rat; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; C000606025:Prom1 protein, rat; D012334:RNA, Neoplasm