OBJECTIVETo probe into the situation and significance of p16 gene CPG island methylation in patients with arseniasis caused by coal-burning pollution.

METHODSDNA was extracted using the Phenol-Chloroform method from leukocytes of 51 patients suffered from coal-burnt arsenism and 52 healthy volunteers. The quantity of the DNA was determined by UV spectrophotometry. Target DNA was denatured by NaOH, then the single strand DNA was modified by sodium bisulfite, converting all unmethylated (but not the methylated) cytosines to uracil. Subsequently a nested amplification with primers specific for methylated versus unmethylated DNA was performed, and PCR products were detected by gel electrophoresis.

RESULTSHypermethylation of the p16 CPG island was presented in 94.1% of the patients suffering from coal-burnt arsenism and in 73.1% of the healthy volunteers. There was statistical difference (P < 0.05) between them.

CONCLUSIONSMethylation of p16 gene CPG island should have important pertinence in the metabolism of coal-burnt arsenism.