Expression of CYP2E1 in human nasopharynx and its metabolic effect in vitro.
It was evident that nitrosamines can act directly on target tissue and result in carcinogenesis. As has been shown, the carcinogenic activity of nitrosamines relied on its bioactivation by Cytochrome P450 2E1 (CYP2E1). In this study, we investigated the expression of CYP2E1 in Nasopharyngeal carcinoma (NPC) cells, embryonic nasopharyngeal epithelial tissue (ENET) specimens, and NPC biopsies by RT-PCR analysis. CYP2E1 was expressed in all NPC cell lines (6/6, including 7429) and ENET (6/6), and 80% of NPC biopsie (8/10). The fact that Human nasopharynx expresses CYP2E1 suggests that CYP2E1 may play an important role in the course of NPC by indirect carcinogens nitrosamines. To further evaluate the function of CYP2E1, the CYP2E1 was stably expressed in the cell line NIH 3T3/rtTA under a tetracycline-controlled transactivator. The expression of CYP2E1 was tightly regulated in a dose-dependent manner by Doxycycline (Dox) When the catalytic activity of CYP2E1 was assayed, the result showed that the generation of 6-hydroxychlorzoxazone (6-OH-CZ) from chlorzoxazone (CZ) was dose- and time-dependent on Dox addition to the medium. In the presence of 1 microg/ml Dox, the CZ 6-hydroxylase activity of the cell line was found to be 0.986 +/- 0.034 nmol/10(6) cells/h. The metabolic activation of Tet/3T3/2E1-6 cells was also assayed by N,N'-dinitrosopiperazine (DNP) cytotoxicity, and the viability of Tet/3T3/2E1-6 cells treated with Dox was lower than that of untreated cells with a significant difference between them in 80 and 160 microg/ml DNP (P ( 0.05, t test. This cell line will be useful not only to assess the metabolic characteristics of CYP2E1, but also will be useful to investigate the role of CYP2E1 in metabolic activation of carcinogenic nitrosamines in vitro.
Hou
De-Fu
DF
Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, 410078, P.R. China.
Wang
Shui-Liang
SL
He
Zhi-Min
ZM
Yang
Fang
F
Chen
Zhu-Chu
ZC
D000818:Animals; D001706:Biopsy; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D002753:Chlorzoxazone / Q000031:analogs & derivatives / Q000378:metabolism; D019392:Cytochrome P-450 CYP2E1 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D009303:Nasopharyngeal Neoplasms / Q000201:enzymology* / Q000473:pathology; D009305:Nasopharynx / Q000187:drug effects / Q000196:embryology / Q000201:enzymology*; D009602:Nitrosamines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013997:Time Factors
D009602:Nitrosamines; D012333:RNA, Messenger; D011994:Recombinant Proteins; C039289:6-hydroxychlorzoxazone; D019392:Cytochrome P-450 CYP2E1; C026156:N,N'-dinitrosopiperazine; D002753:Chlorzoxazone; D004318:Doxycycline