title: In vitro transdifferentiation of HepG2 cells to pancreatic-like cells by CCl₄, D-galactosamine, and ZnCl₂.
abstract: OBJECTIVE
  The objective of the study was to induce transdifferentiation of human hepatoma HepG2 cells into pancreatic-like cells without direct genetic intervention.
  
  
  METHODS
  HepG2 cells were transfected with plasmids for the hepatocyte marker protein green fluorescent protein (albumin-GFP) and the pancreatic cell marker Discosoma spp red fluorescent protein (elastase-DsRed) to create FAE-HepG2 cells. Fluorescent marker expression was used to monitor in vitro transdifferentiation stimulated 100 mM CCl₄, 2 mM D-galactosamine, or 200 μM ZnCl₂. Concentrations were selected for optimal cell survival rate. Transdifferentiation was also characterized by immunohistochemical detection of amylase, glucagon, and insulin and by polymerase change reaction analysis of amylase and insulin mRNA production.
  
  
  RESULTS
  Control cells expressed albumin-GFP but no elastase-DsRed. By 30 days of culture, all 3 agents induced expression of pancreatic-like cell marker elastase-DsRed. ZnCl₂ was the most effective as most cells expressed elastase-DsRed in the absence of simultaneous expression of albumin-GFP. For CCl₄ and D-galactosamine, elastase-DsRed was expressed in the same cells as albumin-GFP. Cells treated by each agent also expressed amylase, insulin, and glucagon proteins and mRNAs.
  
  
  CONCLUSIONS
  Without direct genetic intervention, select low small molecules can induce in vitro transformation of hepatoma cells into pancreatic-like cells.
authors: Department of Histology and Embryology, Shinshu University School of Medicine, Matsumoto, Japan. kanoh@shinshu-u.ac.jp
authors: Yoshiya
authors: Y
authors: Kanoh
authors: Daihachiro
authors: D
authors: Tomotsune
authors: Sakiko
authors: S
authors: Shirasawa
authors: Susumu
authors: S
authors: Yoshie
authors: Hinako
authors: H
authors: Ichikawa
authors: Tadayuki
authors: T
authors: Yokoyama
authors: Shin-Ichi
authors: S
authors: Mae
authors: Jun
authors: J
authors: Ito
authors: Masahiro
authors: M
authors: Mizuguchi
authors: Ken
authors: K
authors: Matsumoto
authors: Fengming
authors: F
authors: Yue
authors: Katsunori
authors: K
authors: Sasaki
mesh_terms: D000418:Albumins / Q000235:genetics / Q000378:metabolism; D000681:Amylases / Q000235:genetics / Q000378:metabolism; D002251:Carbon Tetrachloride / Q000494:pharmacology*; D054338:Cell Transdifferentiation / Q000187:drug effects*; D002712:Chlorides / Q000494:pharmacology*; D005688:Galactosamine / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D005934:Glucagon / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010179:Pancreas / Q000166:cytology* / Q000378:metabolism; D010196:Pancreatic Elastase / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection; D017967:Zinc Compounds / Q000494:pharmacology*
chemical_list: D000418:Albumins; D002712:Chlorides; D007328:Insulin; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D017967:Zinc Compounds; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins; D005688:Galactosamine; C016837:zinc chloride; D005934:Glucagon; D002251:Carbon Tetrachloride; D000681:Amylases; D010196:Pancreatic Elastase