RRC ID 38543
著者 Kanoh Y, Tomotsune D, Shirasawa S, Yoshie S, Ichikawa H, Yokoyama T, Mae S, Ito J, Mizuguchi M, Matsumoto K, Yue F, Sasaki K.
タイトル In vitro transdifferentiation of HepG2 cells to pancreatic-like cells by CCl₄, D-galactosamine, and ZnCl₂.
ジャーナル Pancreas
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.
巻・号 40(8)
ページ 1245-52
公開日 2011-11-1
DOI 10.1097/MPA.0b013e318221933d
PMID 21989025
MeSH Albumins / genetics Albumins / metabolism Amylases / genetics Amylases / metabolism Carbon Tetrachloride / pharmacology* Cell Transdifferentiation / drug effects* Chlorides / pharmacology* Galactosamine / pharmacology* Gene Expression / drug effects Glucagon / genetics Glucagon / metabolism Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Hep G2 Cells Humans Immunohistochemistry Insulin / genetics Insulin / metabolism Luminescent Proteins / genetics Luminescent Proteins / metabolism Microscopy, Fluorescence Pancreas / cytology* Pancreas / metabolism Pancreatic Elastase / genetics Pancreatic Elastase / metabolism Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Reverse Transcriptase Polymerase Chain Reaction Time Factors Transfection Zinc Compounds / pharmacology*
IF 2.92
引用数 4
WOS 分野 GASTROENTEROLOGY & HEPATOLOGY
リソース情報
ヒト・動物細胞