Biochemical and Biophysical Research Communications
Expression of the Musashi1 gene encoding the RNA-binding protein in human hepatoma cell lines☆
Section snippets
Materials and methods
Cell lines and culture. Seven human hepatoma cell lines, HuH6, HuH7, Hep3B, SK-Hep1, HLE, HLF, and HepG2, were used for the mRNA gene-expression analysis of Musashi1. The cells were seeded into 250 ml flasks and maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 2 mM l-glutamine, 100 units/ml penicillin, 100 units/ml streptomycin, and 10% fetal bovine serum at 37 °C in a humidified atmosphere containing 5% CO2. All media and chemicals used in the cell cultures were
Musashi1 protein expression in fetal mouse liver
In mouse fetal liver at 14.5 gestational days, the Musashi1 protein was detected in the cytoplasm of early hepatocytes (Fig. 1).
Musashi1 mRNA expression in human hepatoma cell lines
The PCR-amplified product obtained using the Musashi1-specific primers produced clear bands of the predicted 542-bp size. Musashi1 gene expression was observed in three hepatoma cell lines (HuH6, HuH7, and Hep3B) but was not detected in the other four hepatoma cell lines (SK-Hep1, HepG2, HLE, and HLF) (Fig. 2). Sequencing of the PCR-amplified Musashi1 cDNA in the three
Discussion
The Musashi1 gene, which encodes a neural RNA-binding protein, is closely associated with cell differentiation in CNS stem cells [1], [2], [3], [4], [5]. In the endodermal cells, Musashi1 is known to be expressed in intestinal epithelial stem cells [16]. We demonstrated in this study that, in the mouse, Musashi1 is expressed in early hepatocytes during the embryonic stage of liver development. Abnormalities of cell-regulatory gene expression may be involved in carcinogenesis; however, little is
Acknowledgements
We thank Dr. Hiroshi Kawai (Kanazawa University School of Medicine, Japan), and Dr. Tadahisa Fukui (Yamagata University School of Medicine, Japan) for the kind support of this work.
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This study was supported by a grant-in-aid from the Japan Society for the Promotion of Science (grant number: 13470112). The nucleotide sequence data reported in this paper appear in the DDBJ/EMBL/GenBank nucleotide sequence database with the accession numbers AB072590–AB072592.