293FT is a highly suitable mammalian cell line for the in vitro enzymatic activity analysis of typical P450 proteins.

Mammalian cells have been widely used for the in vitro evaluation of the functional effect of allelic variants of cytochrome P450 (CYP). The aim of this study was to determine the most suitable mammalian cell line for the in vitro drug metabolism analysis of CYP variants. Three reported cell lines (COS-7, HepG2, 293T) and one fast-growing variant of the 293 cell line 293FT were transfected with vectors expressing green fluorescent protein or typical variants of CYP2C9, CYP2C19 or CYP2D6 to investigate the protein expression levels and the catalytic activity of expressed CYP allelic variants. The transfected 293FT cells had the highest protein expression level and exhibited the highest enzymatic activity, while HepG2 cells showed the lowest activity among the four tested cell lines. Simultaneously, 293FT cells still maintained the similar relative enzymatic ratio among three typical CYP2C9 variants to that of the commonly used COS-7 cells. In addition, 293FT cells could also be used for the in vitro functional evaluation of two other typical P450 proteins, CYP2C19 and CYP2D6. Therefore, the 293FT cell line is more suitable for the in vitro enzymatic activity analysis of typical P450 proteins than any other reported mammalian cell lines.

CYP2C9 polymorphism analysis in Han Chinese populations: building the largest allele frequency database.

Genetic polymorphisms of CYP2C9 significantly influence the pharmacokinetics and pharmacodynamics of some drugs, which might result in adverse drug effects and therapeutic failure. Several studies have been performed on CYP2C9 genetic polymorphisms in Han Chinese populations. However, these studies only focused on two commonly investigated alleles, *2 and *3, in relatively small sample sizes. To scale up the gene-scanning region and determine relatively precise data on the genetic distribution pattern in Chinese populations, unrelated healthy Han Chinese volunteers from Zhejiang Province (n=1127) and Hebei (n=1000) Province were recruited as subjects for the direct sequencing of all exons of CYP2C9. As a result, 14 previously reported alleles were detected in this work, and 8 of these alleles (*14, *16, *19, *23, *27, *29, *33 and *34) were described for the first time in Chinese populations. In addition, 37 novel mutations were also detected, of which 22 variants were non-synonymous, and 21 new alleles, *36-*56, were designated by the Human CYP Allele Nomenclature Committee. In vitro functional analysis of these 22 novel CYP2C9 variants revealed that 17 mutations had a significant influence on the protein's catalytic activity. Our study provides the most accurate data on CYP2C9 polymorphisms in Han Chinese populations and detects the largest number of novel allelic variants existing to date. These new alleles will greatly enrich the current knowledge of naturally occurring CYP2C9 variants in Chinese populations.