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荧光原位杂交技术在前列腺腺癌脱氧核糖核酸倍体分析中的应用。

Use of fluorescent in situ hybridization for deoxyribonucleic acid ploidy analysis of prostatic adenocarcinoma.

作者信息

Persons D L, Gibney D J, Katzmann J A, Lieber M M, Farrow G M, Jenkins R B

机构信息

Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.

出版信息

J Urol. 1993 Jul;150(1):120-5. doi: 10.1016/s0022-5347(17)35412-5.

Abstract

Fluorescent in situ hybridization using 2 chromosome specific centromere probes was evaluated as a method of ploidy analysis in touch preparations from 50 radical prostatectomy specimens. Tumors were classified as aneuploid by fluorescent in situ hybridization when nuclei had an abnormal copy number (aneusomic) for either chromosome centromere 8 or 12. Tetraploid tumors were defined as those with 4 copies (tetrasomic) of chromosome centromeres 8 and 12. The fluorescent in situ hybridization ploidy patterns were compared to the deoxyribonucleic acid (DNA) ploidy patterns subsequently obtained by flow cytometry on the same tissue following paraffin embedding. Concordant fluorescent in situ hybridization and flow cytometry ploidy classification was obtained in 82% of the cases (p < or = 0.0001). Of 7 aneuploid tumors 3 were identified by both methods. Trisomy 8 was detected by fluorescent in situ hybridization in 3 cases that were classified as DNA diploid (2 tumors) and DNA tetraploid (1 tumor). Conversely, flow cytometry detected aneuploidy (hypotetraploidy) in 1 tumor when the fluorescent in situ hybridization results were consistent with tetraploidy. Overall, fluorescent in situ hybridization was more sensitive in aneuploidy detection (6 of 7 cases) than flow cytometry (4 of 7). Of 19 tetraploid cases 5 had discordant fluorescent in situ hybridization and flow cytometry results. However, all 5 cases contained low levels of tetraploidy and the discrepant results were most likely due to the limits of precision of 1 or both methods. In conclusion, we demonstrated that fluorescent in situ hybridization ploidy analysis can be rapidly performed on fresh touch preparations of prostate tissue. This preliminary study demonstrates that the ploidy result determined by fluorescent in situ hybridization correlates well with that obtained by flow cytometry. More complete fluorescent in situ hybridization studies of prostate carcinoma will require additional probes for other chromosomes.

摘要

使用2种染色体特异性着丝粒探针的荧光原位杂交技术,被评估为一种对50例根治性前列腺切除术标本的触摸涂片进行倍性分析的方法。当细胞核中8号或12号染色体着丝粒的拷贝数异常(非整倍体)时,荧光原位杂交将肿瘤分类为非整倍体。四倍体肿瘤定义为8号和12号染色体着丝粒有4个拷贝(四体)的肿瘤。将荧光原位杂交的倍性模式与随后在石蜡包埋后的同一组织上通过流式细胞术获得的脱氧核糖核酸(DNA)倍性模式进行比较。82%的病例获得了一致的荧光原位杂交和流式细胞术倍性分类(p≤0.0001)。在7例非整倍体肿瘤中,两种方法均鉴定出3例。荧光原位杂交在3例被分类为DNA二倍体(2例肿瘤)和DNA四倍体(1例肿瘤)的病例中检测到8号染色体三体。相反,当荧光原位杂交结果与四倍体一致时,流式细胞术在1例肿瘤中检测到非整倍体(亚四倍体)。总体而言,荧光原位杂交在非整倍体检测中(7例中的6例)比流式细胞术(7例中的4例)更敏感。在19例四倍体病例中,5例荧光原位杂交和流式细胞术结果不一致。然而,所有5例病例的四倍体水平都很低,差异结果很可能是由于一种或两种方法的精度限制。总之,我们证明荧光原位杂交倍性分析可以在前列腺组织的新鲜触摸涂片上快速进行。这项初步研究表明,荧光原位杂交确定的倍性结果与流式细胞术获得的结果相关性良好。对前列腺癌进行更全面的荧光原位杂交研究将需要针对其他染色体的额外探针。

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