Wu Chang-Xue, Shan Ke-Ren, Zhou Yun-Shu, Yang Qin, Li Cheng-Xiu, Zhao Yan, Cheng Ming-Liang
The Key Laboratory of Molecular Biology, Guiyang Medical College, Guizhou 550004, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2007 Jan;41(1):29-32.
To probe into the situation and significance of p16 gene CPG island methylation in patients with arseniasis caused by coal-burning pollution.
DNA 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.
Hypermethylation 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.
Methylation of p16 gene CPG island should have important pertinence in the metabolism of coal-burnt arsenism.
探讨燃煤污染型砷中毒患者p16基因启动子区甲基化情况及其意义。
采用酚-氯仿法从51例燃煤型砷中毒患者及52名健康对照者外周血白细胞中提取基因组DNA，紫外分光光度计测定DNA含量。用NaOH将模板DNA变性，亚硫酸氢钠修饰单链DNA，使未甲基化的胞嘧啶转变为尿嘧啶，而甲基化的胞嘧啶不变。随后用甲基化特异性引物进行巢式PCR扩增，产物经琼脂糖凝胶电泳分析。
燃煤型砷中毒患者p16基因启动子区甲基化率为94.1%，健康对照者为73.1%，两组比较差异有统计学意义（P < 0.05）。
p16基因启动子区甲基化可能在燃煤型砷中毒发病机制中具有重要作用。
