多色荧光原位杂交检测苯系物接触工人精子1,18号染色体畸变率

Detection of the Frequencies of Numerical and Structural Chromosome Aberrations in Sperm of Benzene Series-exposed Workers by Multi-color Fluorescence in situ Hybridization

为研究苯系物暴露对工人精子染色体的损伤 ,用 4条DNA探针与间期精子核染色体进行多色荧光原位杂交 ,同时检测精子 1号、18号染色体数目畸变和 1号染色体结构畸变 (末端缺失与重复 )。作业车间空气中苯的时间加权浓度 (TWA)为 4 2 2 9mg m3,高于国家卫生标准 (6mg m3)。暴露组工人尿粘糠酸 (ttMA)高于对照组。共计数15例暴露组工人 14 4 2 82条精子 ,14例对照组工人 135 937条精子 ,杂交效率为 99 85 %。非整倍体测定结果 :暴露组精子 1号、18号染色体双体率 (分别为 0 0 88%± 0 0 4 1% ,0 0 87%± 0 0 4 9% )显著高于对照组 1号、18号双体率(0 0 4 5 %± 0 0 2 4 % ,0 0 5 3%± 0 0 2 8% ) ;暴露组 1号、18号染色体缺体率分别为 (0 11%± 0 0 5 9% ,0 0 75 %±0 0 35 % )显著高于对照组相应数值 (0 0 4 8%± 0 0 18% ,0 0 4 5 %± 0 0 2 4 % ) ;而二倍体精子率 ,两组差别无显著性。结构畸变测定结果 :暴露组 1号染色体的末端重复率、末端缺失率 (分别为 0 16 %± 0 0 37% ,0 14 %± 0 0 5 3% )显著高于对照组数值 (分别为 0 0 82 %± 0 0 2 3% ,0 0 6 9%± 0 0 2 8% ) ;暴露组 1号染色体着丝粒重复率及着丝粒缺失率(0 10 %± 0 0 35 % ,0 10 %± 0 0 4 1% )

To study the frequencies of numerical and structural aberrations for chromosome in sperm of benzene exposed workers,the multi color FISH was used.Four DNA probes(one for chromosome 1 centromere and one for 1p terminal,and two for chromosome 18 centromere) were hybridized with interphase sperms,and the frequencies of numerical aberrations for chromosome 1,18 and structural aberrations of chromosome 1 were detected simultaneously.The time weighted average concentration(TWA) of benzene in workplace (42 29 mg/m 3) was higher than that of our national maximum allowable concentration (6 mg/m 3).The geometric concentration of urinary trans,trans muconic acid(tt MA) in exposed group was significantly higher than that of control group.A total of 144 282 sperm of 15 benzene exposed workers and 135 937 sperm in 14 controls were scored.The frequency of hybridization efficiency was 99 85%.The mean frequencies of disomic sperms for chromosome 1 and 18 in exposed group(0 088%±0 041%,and 0 087%±0 049%,respectively) were statistically increased over that of the control group(0 045%±0 024%,and 0 053%±0 028%),and the mean frequencies of nullisomic sperms for chromosome 1 and 18(0 11%±0 059%,0 075%±0 035%) in exposed group were statistically increased over that of control group too(0 048%±0 018%;0 045%±0 024%).The frequencies of diploidy sperm were no difference in both exposed and control groups.The mean frequencies of terminal duplication and terminal deletion for chromosome 1p(0 16%±0 037%;0 14%±0 053%,respectively) were significantly increased over that of control group(0 082%±0 023%;0 069%±0 028%,respectively).The mean frequencies of centromeric duplication and centromeric deletion for chromosome 1(0 10%±0 035%;0 10%±0 041%,respectively) were significantly increased over that of control group(0 075%±0 023%;0 060%±0 029%).Our experiments showed that exposed to benzene at higher concentration(42 29 mg/m 3) may induce increases in frequencies not only of numerical aberrations for chromosome 1 and 18,but also of structural aberrations for chromosome 1 of sperms in exposed workers.