前列腺癌的p16基因甲基化程度定量分析

目的 建立和评价基因组DNA甲基化的定量分析方法,并比较正常前列腺组织和前列腺癌组织中的p16抑癌基因的DNA甲基化差异.方法 提取正常前列腺组织和前列腺癌组织中的基因组DNA,用亚硫酸氢钠将未甲基化的胞嘧啶转化为尿嘧啶,经过PCR扩增后,用BstUⅠ和HpaⅡ限制性内切酶消化,通过电泳分离消化的片段,定量分析基因组中...

Quantitative analysis of p16 gene methylation in prostate tissues

Objective To establish and evaluate a quantitative method for analyzing genomic DNA methylation level and compare p16 tumor suppressor gene methylation levels between normal prostate cells and prostate cancer cells. Methods The genomic DNA was extracted from prostate tissues and converted with sodium bisulfite. The converted DNA was amplified by PCR, and then treated with BstUI and Hpa Ⅱ restriction enzymes. After eleetrophoresis separation, the DNA methylation levels were quantitatively analyzed. Results In normal prostate cells, p16 gene was at low methylation level （about 30% ） , in cancer cells, most samples had a higher methylation level （about 60 -70% ） , while in some samples no remarkable methylation changes were observed. Conclusion As an epigenetic research tool, this method can quantitatively analyze DNA methylation level. The analysis of p16 gene in prostate cancer cells also shows that the methylation level in these cells is higher than in normal cells.