mRNA expression and methylation status of p15 promoter in mouse bone marrow cells exposed to 1,4-benzoquinone / 中华劳动卫生职业病杂志
Chinese Journal of Industrial Hygiene and Occupational Diseases
;
(12): 28-32, 2011.
Article
in Chinese
| WPRIM
| ID: wpr-293764
ABSTRACT
<p><b>OBJECTIVE</b>To detect the expression and the CpG island methylation status of tumor suppressor gene p15 after exposure to 1,4-benzoquinone (1,4-BQ) in primary cultivated C57BL/6J mouse bone marrow cells in vitro.</p><p><b>METHODS</b>The mouse bone marrow cells were isolated in vitro. The effect of 0, 0.1, 1, 5, 10, 20, and 40 µmol/L 1,4-BQ on cell viability (CKK-8) was detected. 0, 0.1, 1, 10 µmol/L 1,4-BQ were used to intoxicate the mouse bone marrow cells for 24 h; Real-time PCR was employed to analyze the mRNA expression level of p15; The bisulfite sequencing PCR (BSP) was used to look into the methylation status of CpG islands in p15 promoter region.</p><p><b>RESULTS</b>1,4-BQ exhibited dose-dependent toxicity to mouse bone marrow cells, and the LC(50) was 8.3 µmol/L (95%CI 4.6 - 10.6 µmol/L). The mRNA expression of p15 in 10 µmol/L group was only equivalent to 43% of control group. Compared with control group, the decrease of p15 mRNA expression in1 and 10 µmol/L concentration were obvious, and the differences had statistical significance (P < 0.05 or P < 0.01). BSP sequencing results were same between the exposure groups and control group, the 56 CpG sites on CpG islands remained in the state of unmethylated.</p><p><b>CONCLUSION</b>mRNA expression of p15 gene decreases after exposure to 1,4-BQ, but the CpG islands methylation status in promoter is not affected, suggesting that methylation does not participate in 1,4-BQ-mediated p15 gene expression decrease, other effect mechanisms still need to be investigated.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
RNA, Messenger
/
Bone Marrow Cells
/
Base Sequence
/
Cells, Cultured
/
Benzoquinones
/
Promoter Regions, Genetic
/
CpG Islands
/
DNA Methylation
/
Environmental Exposure
/
Cyclin-Dependent Kinase Inhibitor p15
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Industrial Hygiene and Occupational Diseases
Year:
2011
Type:
Article
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