Abstract:
AIM:Over 90% of drugs are metabolized by the cytochrome P-450 (CYP) family of liver isoenzymes. The most important enzymes are CYP1A2, 3A4, 2C9/19, 2D6 and 2E1. Although CYP2D6 accounts for <2% of the total CYP liver enzyme content, it mediates metabolism in almost 25% of drugs. In order to study its enzymatic activity for drug metabolism, its cDNA was cloned and a HepG2 cell line stably expressing CYP2D6 was established. METHODS:Human CYP2D6 cDNA was amplified with reverse transcription-polymerase chain reaction (RT-PCR) from total RNA extracted from human liver tissue and cloned into pGEM-T vector. cDNA segment was identified by DNA sequencing and subcloned into a mammalian expression vector pREP9. A cell line was established by transfecting the recombinant plasmid of pREP9-CYP2D6 to hepatoma HepG2 cells. Expression of mRNA was validated by RT-PCR. Enzyme activity of catalyzing dextromethorphan O-demethylation in postmitochondrial supernatant (S9) fraction of the cells was determined by high performance liquid chromatography (HPLC). RESULTS:The cloned cDNA had 4 base differences, e.g. 100 C-T, 336 T-C, 408 C-G and 1 457 G-C, which resulted in P34S, and S486T amino acid substitutions, and two samesense mutations were 112 F and 136 V compared with that reported by Kimura et al (GenBank accession number: M33388). P34S and S486T amino acid substitutions were the characteristics of CYP2D6*10 allele. The relative activity of S9 fraction of HepG2-CYP2D6*10 metabolized detromethorphan O-demethylation was found to be 2.31 +/- 0.19 nmol/min(-1)/mg(-1) S9 protein (n=3), but was undetectable in parental HepG2 cells. CONCLUSION:cDNA of human CYP2D6*10 can be successfully cloned. A cell line, HepG2-CYP2D6*10, expressing CYP2D6*10 mRNA and having metabolic activity, has been established.
journal_name
World J Gastroenteroljournal_title
World journal of gastroenterologyauthors
Zhuge J,Yu YN,Wu XDdoi
10.3748/wjg.v10.i2.234keywords:
subject
Has Abstractpub_date
2004-01-15 00:00:00pages
234-7issue
2eissn
1007-9327issn
2219-2840journal_volume
10pub_type
杂志文章abstract::The aetiology of primary sclerosing cholangitis (PSC) is not known and controversy exists as to whether PSC should be denominated an autoimmune disease. A large number of autoantibodies have been detected in PSC patients, but the specificity of these antibodies is generally low, and the frequencies vary largely betwee...
journal_title:World journal of gastroenterology
pub_type: 社论,评审
doi:10.3748/wjg.14.3781
更新日期:2008-06-28 00:00:00
abstract:AIM:To evaluate whether FDG-positron emission tomography (PET)/computed tomography (CT) may be an accurate technique in the assessment of the T stage in patients with colorectal cancer. METHODS:Thirty four consecutive patients (20 men and 14 women; mean age: 63 years) with a histologically proven diagnosis of colorect...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v17.i11.1427
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v13.i32.4295
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abstract::Studies on rodents and humans demonstrate an inherited predisposition to hepatocellular carcinoma (HCC). Analysis of the molecular alterations involved in the acquisition of a phenotype resistant or susceptible to hepatocarcinogenesis showed a deregulation of G1 and S phases in HCC of genetically susceptible F344 rats...
journal_title:World journal of gastroenterology
pub_type: 社论
doi:10.3748/wjg.14.6601
更新日期:2008-11-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v9.i10.2382
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abstract:AIM:To summarize progress in the study of K-ras gene studies in pancreatic cancer and its potential clinical significance in screening test for early detection of pancreatic cancer, and to differentiate pancreatic cancer from chronic pancreatitis in recent decade. METHODS:Literature search (MEDLINE 1986-2003) was perf...
journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v10.i4.471
更新日期:2004-02-15 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v16.i14.1765
更新日期:2010-04-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v21.i26.8089
更新日期:2015-07-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v9.i2.193
更新日期:2003-02-01 00:00:00
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pub_type: 杂志文章,多中心研究
doi:10.3748/wjg.v20.i34.12260
更新日期:2014-09-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v21.i30.9021
更新日期:2015-08-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v23.i7.1203
更新日期:2017-02-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 临床试验,杂志文章
doi:10.3748/wjg.v23.i17.3133
更新日期:2017-05-07 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v19.i27.4380
更新日期:2013-07-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v26.i17.1993
更新日期:2020-05-07 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v16.i23.2851
更新日期:2010-06-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v10.i19.2911
更新日期:2004-10-01 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 临床试验,杂志文章,随机对照试验
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v13.i44.5938
更新日期:2007-11-28 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 临床试验,杂志文章
doi:10.3748/wjg.v11.i8.1109
更新日期:2005-02-28 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v18.i10.1059
更新日期:2012-03-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v21.i3.953
更新日期:2015-01-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,评审
doi:10.3748/wjg.v17.i27.3213
更新日期:2011-07-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v12.i33.5408
更新日期:2006-09-07 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v13.i3.462
更新日期:2007-01-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章,多中心研究
doi:10.3748/wjg.14.6195
更新日期:2008-10-28 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 评论,信件
doi:10.3748/wjg.14.2617
更新日期:2008-04-28 00:00:00
abstract:AIM:To elucidate the sequential gene expression profile in AGS cells co-cultured with wild-type Helicobacter pylori (H pylori) as a model of H pylori-infected gastric epithelium, and to further examine the contribution of cag-pathogenicity islands (cagPAI)-coding type IV secretion system and the two pathways, nuclear f...
journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i39.6134
更新日期:2005-10-21 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v19.i38.6416
更新日期:2013-10-14 00:00:00
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journal_title:World journal of gastroenterology
pub_type: 杂志文章
doi:10.3748/wjg.v11.i41.6422
更新日期:2005-11-07 00:00:00