焦炉工人周围血细胞p16基因甲基化研究

目的分析焦炉工人周围血细胞中p16基因启动子区CpG岛异常甲基化状态,寻找焦炉工人肺癌辅助筛查的生物标志物。方法以74名男性焦炉工人为接触组,以47名供水厂男性职工为对照组。采集2组人员的班后尿,用高效液相色谱法检测其内暴露指标1-羟基芘(1-OH-Py)的水平;同时采集晨起空腹肘静脉血,分离周围血单个核细胞,用彗星实验检测DNA损伤情况;并提取基因组DNA,用甲基化特异性聚合酶链反应技术检测p16基因启动子区CpG岛甲基化发生情况。结果接触组尿1-OH-Py水平(0.46±0.12)μmol/mol Cr]和DNA拖尾的Olive尾距(0.41±0.25)μm]高于对照组(0.17±0.06)μmol/mol Cr、(0.32±0.12)μm],差异有统计学意义(P<0.05)。接触组p16基因的异常甲基化的检出率(36.49%)高于对照组(4.26%),差异有统计学意义(P<0.01);并且p16基因的异常甲基化检出率随着尿1-OH-Py的水平升高逐渐增加,差异有统计学意义(P<0.01)。结论周围血p16基因异常甲基化可能是焦炉工人肺癌早期筛查有效的生物标志物。

Study on p16 gene methylation in peripheral blood cells of coke oven workers

Objective To analyze p16 gene methylation in the promoter CpG island,and to find the biomarker for screening the lung cancer of the coke oven workers.Methods The subjects were consisted of 74 male coke oven workers(the exposed group) and 47 male workers employed in a water supply plant(the controls).Post-shift urine samples in 2 groups were collected and 1-hydroxypyrene levels were detected by high performance liquid chromatography.Venous blood was drawn in the fast mor-ning and peripheral blood mononuclear cells were separated for DNA damage determination by comet assay.Genomic DNA was extracted to detect p16 gene methylation in the promoter CpG island with methylation specific polymerase chain reaction(MS-PCR) methods.Results Urinary 1-OH-Py level in the exposed group(0.46±0.12) μmol/mol Cr and DNA Olive Tail Moment(OTM)(0.41±0.25)μm were significantly higher than those of the control group 1-OH-Py level(0.17±0.06) μmol/mol Cr and OTM(0.32±0.12) μm].The frequency of p16 gene methylation in the coke oven workers(36.49%) were significantly higher than that in the control group(4.26%),with the increase of 1-OH-Py level in the exposed group.Conclusion p16 gene methylation may be the biomarker for lung cancer screening in the coke oven workers.