Identification of Dioxin-Responsive Genes in Hep G2 Cells Using Differential mRNA Display RT–PCR

• Published in: Biochemical and biophysical research communications 1996-03, Vol.220 (3), p.784-788
• Main Authors: Wang, Xiaohong, Harris, Peter K.W., Ulrich, Roger G., Voorman, Richard L.
• Format: Article
• Language: English
• Subjects: Base Sequence; Blotting, Northern; Carcinoma, Hepatocellular; Cell Line; Cloning, Molecular; Cytochrome P-450 Enzyme System - biosynthesis; Databases, Factual; DNA Primers; DNA, Complementary; Gene Expression Regulation, Neoplastic - drug effects; Humans; Liver Neoplasms; Molecular Sequence Data; Polychlorinated Dibenzodioxins - pharmacology; Polymerase Chain Reaction - methods; RNA, Messenger - biosynthesis; Sequence Homology, Nucleic Acid; Software; Tumor Cells, Cultured
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1996.0481

Differential mRNA display RT–PCR (DD RT–PCR) offers a tool to identify genes which are regulated or responsive to certain receptors or chemicals such as dioxin (TCDD). Treatment of Hep G2 cells with TCDD followed by DD analysis of a gel with series of different primers revealed a significantly different pattern from the control for a number of mRNAs. The differentially displayed mRNAs were isolated and reamplified. A GenBank search of four mRNAs revealed two known and two unknown sequences. Northern blot analysis revealed that two known sequences, fibrinogen γ chain and plastin mRNAs were down regulated by TCDD in a time-dependent manner, whereas two unknown mRNAs were induced by TCDD treatment. The function of these genes in TCDD toxicity is not known; however, the application of DD RT–PCR in the studies of TCDD-induced responses could be very useful in the discovery of other unknown genes important for TCDD toxicity.