Author(s): LeCluyse EL
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Abstract Primary cultures of human hepatocytes have been used extensively by both academic and industrial laboratories for evaluating the hepatic disposition of drugs and other xenobiotics. Their primary utility has been for assessing the induction potential of new chemical entities (NCEs) and they continue to serve as the gold standard. Primary considerations for conducting in vitro drug testing utilizing cultures of human hepatocytes, such as the effects of culture and study conditions, are discussed. The maintenance of normal cellular physiology and intercellular contacts in vitro is of particular importance for optimal phenotypic gene expression and response to drugs and other xenobiotics. Significant advances in our understanding of cytochrome P450 (CYP450) enzyme regulation have been made with the recent identification of the nuclear receptors mediating the induction of CYP2B and CYP3A enzymes. In particular, the activation of pregnane X receptor (PXR) by prototypical inducers of CYP3A has been found to correlate well with the species-specific modulation of CYP3A by various drugs and other xenobiotics. Concomitant with the discovery of PXR has been the identification of compounds that may act synergistically or antagonistically on multiple receptors (e.g., co-repressors and/or co-activators of the receptor) introducing novel mechanisms of drug-drug interactions. Differential expression of the individual isoforms of the major CYP450 enzymes over time in culture suggest that this model system is not reflective of in vivo profiles and, therefore, may be limited in its application for drug metabolism studies. Overall, primary cultures of human hepatocytes can serve as a sensitive and selective model for predicting the regulation of CYP450 modulation by drugs and other xenobiotics. Considerations and recommendations for standardizing testing conditions and choosing relevant endpoint(s) are presented.
This article was published in Eur J Pharm Sci
and referenced in Journal of Bioengineering & Biomedical Science
- Yosef Yarden
Classically, the 3âuntranslated region (3âUTR) is that region in eukaryotic protein-coding genes from the translation termination codon to the polyA signal. It is transcribed as an integral part of the mRNA encoded by the gene. However, there exists another kind of RNA, which consists of the 3âUTR alone, without all other elements in mRNA such as 5âUTR and coding region. The importance of independent 3âUTR RNA (referred as I3âUTR) was prompted by results of artificially introducing such RNA species into malignant mammalian cells. Since 1991, we found that the middle part of the 3âUTR of the human nuclear factor for interleukin-6 (NF-IL6) or C/EBP gene exerted tumor suppression effect in vivo. Our subsequent studies showed that transfection of C/EBP 3âUTR led to down-regulation of several genes favorable for malignancy and to up-regulation of some genes favorable for phenotypic reversion. Also, it was shown that the sequences near the termini of the C/EBP 3âUTR were important for its tumor suppression activity. Then, the C/EBP 3âUTR was found to directly inhibit the phosphorylation activity of protein kinase CPKC in SMMC-7721, a hepatocarcinoma cell line. Recently, an AU-rich region in the C/EBP 3âUTR was found also to be responsible for its tumor suppression. Recently we have also found evidence that the independent C/EBP 3âUTR RNA is actually exists in human tissues, such as fetal liver and heart, pregnant uterus, senescent fibroblasts etc. Through 1990âs to 2000âs, world scientists found several 3âUTR RNAs that functioned as artificial independent RNAs in cancer cells and resulted in tumor suppression. Interestingly, majority of genes for these RNAs have promoter-like structures in their 3âUTR regions, although the existence of their transcribed products as independent 3âUTR RNAs is still to be confirmed. Our studies indicate that the independent 3âUTR RNA is a novel non-coding RNA species whose function should be the regulation not of the expression of their original mRNA, but of some essential life activities of the cell as a whole.
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- Abel E Navarro
Improving the adsorptive properties of biomaterials for the removal of heavy metals
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