Quantification of cyclooxygenase-2 methylation in gastric mucosa by denaturing high performance liquid chromatography assay / 中华预防医学杂志
Chinese Journal of Preventive Medicine
; (12): 54-57, 2010.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-291561
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To setup a quantitative assay for detection of cyclooxygenase-2 (COX-2) methylation in human gastric mucosa samples.</p><p><b>METHODS</b>A standard analysis system was established by denaturing high performance liquid chromatography (DHPLC) under the condition of 55 degrees C oven temperature and a linear acetonitrile gradient (4.0/min). While, a total of 10 cases of gastric biopsy samples were detected for methylation status of COX-2.</p><p><b>RESULTS</b>The complete methylated human promyelocytic leukemia cells (HL-60) and unmethylated gastric cancer cell line (MGC803) were used as positive and negative control. The proportion of the methylated copies of COX-2 was calculated according to the peak heights of methylated (M) and unmethylated (U) COX-2 in same PCR amplicon. The formula was Y = 1.0608 x M/(M + U), R(2) = 0.9894. Among 10 biopsy samples, the proportions of methylated copies of COX-2 in 2 cases of dysplasia were higher than superficial gastritis and chronic atrophy gastritis (24.5%, 18.4% vs 7.6%, 9.6%).</p><p><b>CONCLUSION</b>The methylation of COX-2 promoter CpG islands can be detected in human gastric mucosa samples by quantitative DHPLC assay, which could be used in the population-based study of precancerous gastric lesions.</p>
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Stomach Diseases
/
Chromatography, High Pressure Liquid
/
Promoter Regions, Genetic
/
CpG Islands
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DNA Methylation
/
Cell Line, Tumor
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Cyclooxygenase 2
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Gastric Mucosa
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Genetics
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Preventive Medicine
Year:
2010
Document type:
Article