用间期荧光原位杂交检测卵巢癌细胞中X染色体数目的异常

Application of Interphase Fluorescence in situ Hybridization to the Diagnosis of X Chromosomal Count Abnormality in Ovarian Carcinoma Cell

为了探讨用荧光原位杂交技术(fluorescence in situ hybridization, FISH)检测卵巢癌细胞中性染色体拷贝数目异常的实验方法及其应用价值,收集18例新鲜卵巢癌组织标本,以Biotin标记的X染色体α 卫星DNA(pBamX7)探针与经处理的标本进行卵巢癌细胞核的原位杂交,分别用 Avidin FITC和 Anti avidin进行信号的检测与放大,PI复染。于Olympus AX 70型荧光显微镜下,通过WIB滤光镜观察杂交信号及其细胞核背景,并统计卵巢癌细胞核中的杂交信号颗粒数量。在显微镜下可见以Biotin标记的 pBamX7探针显示绿色杂交信号,细胞核背景经 PI复染显示桔红色;发现11/18(61%)卵巢癌标本中X染色体拷贝数增加,其余7例(39%)无拷贝数增加。X染色体拷贝数目增多在卵巢癌中有一定比例的发生频率,其在促进卵巢癌发病及其发展过程中起到某种作用,其意义值得进一步研究。

To study the technique of fluorescence in situ hybridization (FISH) and its application in the diagnosis of sex chromosomal count abnormality in ovarian carcinoma cell, biotin labeled alpha satellite X chromosome DNA(pBamX7) probe was hybridized with pre-treated slides of ovarian carcinoma cell interphase nucleus in 18 cases of ovarian carcinoma specimens. The slides were treated with Avidin-FITC and Anti-avidin, amplified with an additional layer and counter-stained with PI in antifade solution. The hybridization signals as well as interphase nucleus settings were observed with WIB filters under fluorescence microscope Olympus AX-70, and the number of interphase nucleus in the ovarian carcinoma cell was counted. It was observed under the microscope that the biotin labeled pBamX7 probe showed green hybridization signals, and cytoplasm counter-stained with PI showed reddish orange. Increased chromosome X copy number was observed in 11/18(61%) ovarian carcinoma specimens, while the rest 7 (39%) had no increase in chromosome X copy number. Gain of X chromosome had a certain incidence in ovarian cancers, which played a role in the recurrence and development of ovarian cancers. Its significance needs further investigation.