Decreased expression of glutathione S-transferase M1 in HPV16-transfected human cervical keratinocytes in culture.
Glutathione S-transferase (GST) M1 is a member of the GST mu family of cytosolic enzymes that have been hypothesized to catalyze the conjugation of glutathione to a large number of hydrophobic substances, including carcinogens such as polynuclear aromatic hydrocarbons present in tobacco smoke, leading to their excretion. Epidemiologic and experimental evidence suggests that the risk of cervical cancer is related to both human papillomavirus (HPV) infection and cigarette smoking. We compared the enzymatic activities and mRNA levels of GSTs in GSTM1-positive human cervical keratinocytes (HCKs) that had been transfected with HPV16 with those in the parental cells. The GSTM1 activity toward the substrate trans-stilbene oxide was 5- to 7-fold lower than in the parental cells. The relative mRNA level in HCK transfected with HPV16 E6/E7, as quantified by reverse transcriptase-polymerase chain reaction (RT-PCR) with normalization against endogenous glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression, was 6% that of the parental cells. It was 16 and 82%, respectively, in cells that were transfected with HPV16 E6 alone or HPV16 E7 alone. When quantified by competitive RT-PCR using an exogenous nuclease-resistant synthetic cyclophilin RNA transcript as control, the mRNA level in HCK transfected with HPV16 E6 was approximately 10-fold lower that that in the parental cells. It was approximately 5- to 7-fold lower in the HPV16 E7 or HPV16 E6/E7 cells. Our results suggest that viral infections, through the modulation of cellular xenobiotic-metabolizing enzymes, may play a role in the ability of cells to handle environmental carcinogens.
Chen
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C
Program in Cancer Biology, Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024, USA. cchen@fhcrc.org
Nirunsuksiri
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W
D002274:Carcinogens, Environmental / Q000493:pharmacokinetics; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000166:cytology; D004198:Disease Susceptibility; D004790:Enzyme Induction; D005260:Female; D005786:Gene Expression Regulation; D015967:Gene Expression Regulation, Viral; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000172:deficiency / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000172:deficiency / Q000235:genetics; D015603:Keratinocytes / Q000201:enzymology* / Q000821:virology; D008297:Male; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000502:physiology*; D050725:Papillomavirus E7 Proteins; D010413:Penis / Q000166:cytology; D012333:RNA, Messenger / Q000096:biosynthesis; D012097:Repressor Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013267:Stilbenes / Q000378:metabolism; D014162:Transfection
D002274:Carcinogens, Environmental; D005982:Glutathione Transferase; D007527:Isoenzymes; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D013267:Stilbenes; C068651:E6 protein, Human papillomavirus type 16; C059731:oncogene protein E7, Human papillomavirus type 16; C025906:stilbene oxide