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使用单拷贝P1探针通过荧光原位杂交在石蜡包埋的前列腺肿瘤分离细胞核上检测到8号、10号和17号染色体上的等位基因缺失。

Allelic loss detected on chromosomes 8, 10, and 17 by fluorescence in situ hybridization using single-copy P1 probes on isolated nuclei from paraffin-embedded prostate tumors.

作者信息

Deubler D A, Williams B J, Zhu X L, Steele M R, Rohr L R, Jensen J C, Stephenson R A, Changus J E, Miller G J, Becich M J, Brothman A R

机构信息

Department of Pediatrics, University of Utah Health Sciences Center, Salt Lake City 84132, USA.

出版信息

Am J Pathol. 1997 Mar;150(3):841-50.

Abstract

We have implemented a reliable new technique for preparing isolated prostate cancer nuclei from paraffin-embedded tissue sections followed by analysis with single-copy fluorescence in situ hybridization (FISH). Our initial validation is described by comparison of our data with fresh prostate tumor tissue and loss of heterozygosity (LOH) studies. We also describe evaluation of 36 previously unstudied prostate cancer patients. Fifteen archival samples were selected from patients who underwent radical prostatectomy in which direct FISH and LOH data were available. Isolated nuclei were prepared and allelic loss was detected on 17q using a single-copy DNA (P1 phage) probe by FISH. A high (80%) concordance was found when comparing isolated nuclei data with 17q results from fresh preparations and LOH studies. We also examined loss at sites on 8p, 10q, and 17q in samples from 36 patients for whom clinical information was available. Loss was found at any of the three loci in 32/36 (89%) of the specimens with specific loss in 53% of the cases at the 8p locus, 33% at the 10q locus, and 61% at the 17q locus. Studies indicate that, as well as providing potential clinical information, isolated nuclei preparations are as reliable as fresh tissue for single-copy FISH studies.

摘要

我们已经实现了一种可靠的新技术,用于从石蜡包埋的组织切片中制备分离的前列腺癌细胞核,随后通过单拷贝荧光原位杂交(FISH)进行分析。我们通过将数据与新鲜前列腺肿瘤组织以及杂合性缺失(LOH)研究进行比较来描述我们的初步验证。我们还描述了对36例先前未研究过的前列腺癌患者的评估。从接受根治性前列腺切除术的患者中选取了15份存档样本,这些样本可获得直接的FISH和LOH数据。制备分离的细胞核,并通过FISH使用单拷贝DNA(P1噬菌体)探针检测17号染色体长臂上的等位基因缺失。将分离细胞核的数据与新鲜样本的17号染色体长臂结果以及LOH研究结果进行比较时,发现高度一致性(80%)。我们还检查了36例有临床信息的患者样本中8号染色体短臂、10号染色体长臂和17号染色体长臂位点的缺失情况。在32/36(89%)的标本中发现三个位点中的任何一个存在缺失,其中8号染色体短臂位点特异性缺失的病例占53%,10号染色体长臂位点占33%,17号染色体长臂位点占61%。研究表明,分离细胞核制备不仅能提供潜在的临床信息,而且对于单拷贝FISH研究而言,其可靠性与新鲜组织相当。

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