Fauth E, Zankl H
Abt. Humanbiologie und Humangenetik der Universität, Postfach 3049, D-67653, Kaiserslautern, Germany.
Mutat Res. 1999 Apr 6;440(2):147-56. doi: 10.1016/s1383-5718(99)00021-2.
Fluorescence in situ hybridisation (FISH) technique with chromosome specific library (CSL) DNA probes for all human chromosomes were used to study about 9000 micronuclei (MN) in normal and idoxuridine (IUdR)-treated lymphocyte cultures of female and male donors. In addition, MN rates and structural chromosome aberrations were scored in Giemsa-stained chromosome spreads of these cultures. IUdR treatment (40 microg/ml) induced on the average a 12-fold increase of the MN rate. Metaphase analysis revealed no distinct increase of chromosome breaks but a preferential decondensation at chromosome 9q12 (28-79%) and to a lower extend at 1q12 (8-21%). Application of FISH technique with CSL probes to one male and one female untreated proband showed that all human chromosomes except chromosome 12 (and to a striking high frequency chromosomes 9, X and Y) occurred in spontaneous MN. In cultures containing IUdR, the chromosomal spectrum found in MN was reduced to 10 chromosomes in the male and 13 in the female proband. Eight chromosomes (2, 6, 12, 13, 14, 15, 17 and 18) did not occur in MN of both probands. On the contrary chromosomes 1 and especially 9 were found much more frequently in the MN of IUdR-treated cultures than in MN of control cultures. DAPI-staining revealed heterochromatin signals in most of the IUdR-induced MN. In an additional study, spontaneous and IUdR-induced MN were investigated in lymphocytes of another female donor using CSL probes only for chromosomes 1, 6, 9, 15, 16 and X. The results confirmed the previous finding that chromosomes 1 and 9 occur very often in MN after IUdR-treatment. The results indicate that decondensation of heterochromatic regions on chromosomes 1 and 9 caused by IUdR treatment strongly correlates with MN formation by these chromosomes.
采用针对所有人类染色体的染色体特异性文库(CSL)DNA探针的荧光原位杂交(FISH)技术,对来自男性和女性供体的正常及经碘苷(IUdR)处理的淋巴细胞培养物中的约9000个微核(MN)进行了研究。此外,对这些培养物经吉姆萨染色的染色体铺片进行了微核率和染色体结构畸变评分。IUdR处理(40微克/毫升)平均使微核率增加了12倍。中期分析显示染色体断裂没有明显增加,但在9号染色体q12区域(28 - 79%)出现了优先解聚,在1号染色体q12区域解聚程度较低(8 - 21%)。对一名未处理的男性和一名未处理的女性先证者应用CSL探针的FISH技术显示,除12号染色体外(以及9号、X和Y染色体出现频率极高),所有人类染色体都出现在自发微核中。在含有IUdR的培养物中,男性先证者微核中发现的染色体谱减少到10条染色体,女性先证者减少到13条染色体。在两名先证者的微核中均未出现8条染色体(2、6、12、13、14、15、17和18号)。相反,在经IUdR处理的培养物的微核中,1号染色体尤其是9号染色体出现的频率比对照培养物的微核中高得多。DAPI染色显示,在大多数IUdR诱导的微核中存在异染色质信号。在另一项研究中,仅使用针对1、6、9、15、16和X号染色体的CSL探针,对另一名女性供体的淋巴细胞中的自发和IUdR诱导的微核进行了研究。结果证实了先前的发现,即IUdR处理后1号和9号染色体经常出现在微核中。结果表明,IUdR处理导致的1号和9号染色体异染色质区域解聚与这些染色体形成微核密切相关。