The influence of culture medium composition on drug metabolising enzyme activities of the human liver derived Hep G2 cell line

Doostdar, Hamed; Duthie, Susan J.; Burke, Mary; Melvin, William T.; Grant, M.H.

doi:10.1016/0014-5793(88)81021-4

Cited by 74 publications

(24 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The ability of each cell type to respond to HAH in serum-free medium should be determined, since serum withdrawal greatly reduces AHR content in Swiss 3T3 cells (Vaziri et al 1996) and can abrogate CYP1A induction in PLHC-1 cells after 48 hours (Chapter 5). As noted previously, the presence of serum also affects the levels of cytochromes P450 in some cultured cells (Doostdar et al 1988;Turner et al 1989;Doostdar et al 1991;Hammond et al 1992). …”

Section: One Solution To This Potential Problem Is To Treat Differentsupporting

confidence: 53%

“…For example, the presence of 10% fetal calf serum reduces the potency of TCDD and PCB126 for inhibiting aromatase (CYPI9) activity in JEG-3 human choriocarcinoma cells (Drenth et al 1998). Serum and other medium components can also alter the detectable levels of cytochromes P450 in rat hepatocytes (Turner and Pitot 1989;Hammond and Fry 1992) and HepG2 cells (Doostdar et al 1988;Doostdar et al 1991). Despite these provocative findings, there has been no quantitative study of the effect of serum on CYP1A induction by HAR.…”

Section: Ahr Cloning and Expressionmentioning

confidence: 99%

See 1 more Smart Citation

Mechanisms of action for aryl hydrocarbon receptor ligands in the PLHC-1 cell line

Hestermann¹

1999

View full text Add to dashboard Cite

Section: One Solution To This Potential Problem Is To Treat Differentsupporting

confidence: 53%

Section: Ahr Cloning and Expressionmentioning

confidence: 99%

Mechanisms of action for aryl hydrocarbon receptor ligands in the PLHC-1 cell line

Hestermann¹

1999

View full text Add to dashboard Cite

“…In addition, cell culture environments, such as the composition of the culture medium and the oxygen concentration, can alter DMET expression profiles in HepG2 cells. Higher expression levels of CYP1A and CYP2B were found in cells cultured in Earle's medium compared with those in Dulbecco's modified Eagle's medium and Williams' E medium (Doostdar et al, 1988). During exposure to moderate hypoxia for 24 h, HepG2, Hep3B, and Huh7 produced a general pattern of down-regulation of response genes including drug-metabolizing genes (Fink et al, 2001).…”

Section: Discussionmentioning

confidence: 93%

Similarities and Differences in the Expression of Drug-Metabolizing Enzymes between Human Hepatic Cell Lines and Primary Human Hepatocytes

Guo

Dial²,

Shi³

et al. 2010

Drug Metab Dispos

285

202

View full text Add to dashboard Cite

ABSTRACT:In addition to primary human hepatocytes, hepatoma cell lines, and transfected nonhepatoma, hepatic cell lines have been used for pharmacological and toxicological studies. However, a systematic evaluation and a general report of the gene expression spectra of drug-metabolizing enzymes and transporters (DMETs) in these in vitro systems are not currently available. To fill this information gap and to provide references for future studies, we systematically characterized the basal gene expression profiles of 251 drugmetabolizing enzymes in untreated primary human hepatocytes from six donors, four commonly used hepatoma cell lines (HepG2, Huh7, SK-Hep-1, and Hep3B), and one transfected human liver epithelial cell line. A large variation in DMET expression spectra was observed between hepatic cell lines and primary hepatocytes, with the complete absence or much lower abundance of certain DMETs in hepatic cell lines. Furthermore, the basal DMET expression spectra of five hepatic cell lines are summarized, providing references for researchers to choose carefully appropriate in vitro models for their studies of drug metabolism and toxicity, especially for studies with drugs in which toxicities are mediated through the formation of reactive metabolites.

show abstract

“…[19][20][21] Although APAP-induced hepatotoxicity is mediated by the reactive electrophilic metabolite NAPQI, converted from APAP by CYP, 4) the expression levels of CYP in HepG2 cells are very low compared with those in normal hepatocytes, 22) suggesting that the reactive intermediate would not be expected to appear in APAP-treated HepG2 cells. Therefore, the altered expression of identified genes must have resulted from the direct effect of APAP itself, not from the cytotoxicity of its metabolites.…”

Section: Discussionmentioning

confidence: 99%

Identification of Differentially Expressed Genes in Hepatic HepG2 Cells Treated with Acetaminophen Using Suppression Subtractive Hybridization

Iguchi

Takahashi

Kaneto

et al. 2005

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

Acetaminophen (N-acetyl-p-aminophenol; paracetamol; APAP) is a widely used drug for the treatment of pain and fever, but overdose produces severe hepatotoxicity in humans and experimental animals.1,2) APAP hepatotoxicity is due to its biotransformation to a reactive metabolite, N-acetyl-pbenzoquinone imine (NAPQI) by cytochrome P450 (CYP), leading to cellular glutathione depletion by NAPQI conjugation followed by covalent binding of NAPQI to cellular proteins.3,4) APAP toxicity was not observed in CYP1A2 and CYP2E1 double-null mice, indicating that the formation of the metabolites of APAP by the CYPs is a critical step in the initiation phase of the hepatotoxicity. 5) Thus, it is indisputable that the reactive metabolite of APAP plays an essential role for the initiation phase of APAP-induced hepatotoxicity. However, in the progression phase following initiation, there is almost no information besides the fact that chemokines are associated with hepatocytes regeneration following toxicity. 6)Since drug metabolism is impaired in patients with liver disease, the total amount of non-metabolized drug in the serum is increased in the patients with damaged liver. 7,8) In the case of hepatotoxicity induced by APAP, high level of non-metabolized APAP in the serum of the patients has been observed. 9) Therefore, evaluation of the effect of non-metabolized APAP on hepatocytes is required for full understanding of the progression phase of APAP-induced hepatotoxicity.There are several studies on the gene expression profiles associated with hepatotoxicity in vitro and in vivo. [10][11][12] These reports reveal the specific gene expression profiles from several types of hepatotoxins using microarray technology. The DNA microarray is a useful technology that was developed as a way to simultaneously evaluate the relative levels of large numbers of mRNA gene transcripts. However, the technology is limited by its insensitivity to transcripts of low abundance.13) In another experimental approach, suppression subtractive hybridization (SSH) was also employed to identify differentially regulated genes. This method makes it possible to detect some low abundance transcripts because of a normalization step to equalize the abundance of target in the subtracted cDNA population. 14,15) In the present study, to clarify the effect of APAP on hepatic cells, we investigated the gene increased in APAPtreated HepG2 cells, which show low metabolic activity for APAP. The identification of the differentially expressed genes was performed with the SSH technique and two genes that had changed their expression levels in APAP-treated HepG2 cells were determined. MATERIALS AND METHODSMaterials APAP was purchased from Merck Hoei (Osaka, Japan). Polymerase chain reaction (PCR) primers were purchased from Sigma Genosys (Ishikari, Japan). All other chemicals were of analytical grade or equivalent, and purchased commercially.Cell Culture Human hepatic HepG2 cells were purchased from American Type Tissue Collection (ATCC, Rockvill, MD, U.S.A.), and cultured ...

show abstract

The influence of culture medium composition on drug metabolising enzyme activities of the human liver derived Hep G2 cell line

Cited by 74 publications

References 25 publications

Mechanisms of action for aryl hydrocarbon receptor ligands in the PLHC-1 cell line

Mechanisms of action for aryl hydrocarbon receptor ligands in the PLHC-1 cell line

Similarities and Differences in the Expression of Drug-Metabolizing Enzymes between Human Hepatic Cell Lines and Primary Human Hepatocytes

Identification of Differentially Expressed Genes in Hepatic HepG2 Cells Treated with Acetaminophen Using Suppression Subtractive Hybridization

Contact Info

Product

Resources

About