新一代前列腺癌生物银行:面向国际癌症基因组联盟的处理方案。

Raquel Esgueva, Kyung Park, Robert Kim, Naoki Kitabayashi, Christopher E Barbieri, Philip J Dorsey, Cyril Abraham, Samprit Banerjee, Robert A Leung, Ashutosh K Tewari, Stéphane Terry, Maria M Shevchuk, David S Rickman, Mark A Rubin
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引用次数: 28

摘要

下一代DNA和RNA测序需要来自高质量人体组织样本的完整核酸,以更好地阐明癌症的分子基础。我们已经开发了一种前列腺生物银行方案,以获得合适的样本进行测序,而不影响临床诊断的准确性。为了评估实施该方案的临床意义,我们评估了2008年11月至2009年2月连续105例根治性前列腺切除术标本。交替水平的前列腺样本作为福尔马林固定、石蜡包埋块提交外科病理学,作为冷冻块提交机构生物库。比较了临床和获得标本之间报告的病理特征的差异。临床分期和分级不受生物库方案的影响。鉴定冷冻苏木精和伊红载玻片上的肿瘤病灶,对高密度肿瘤病灶进行评分,提取DNA和RNA进行测序。从22例44例高密度肿瘤灶中提取DNA和RNA。82%(18/22)的样本通过了DNA和RNA测序的严格质量控制步骤。迄今为止,已有7例提取的DNA进行了全基因组测序,18例提取的RNA进行了RNA测序。该方案为高通量生物医学研究提供了前列腺组织,并证实了积极将前列腺癌纳入国际癌症基因组联盟成员癌症基因组图谱计划的可行性。
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Next-generation prostate cancer biobanking: toward a processing protocol amenable for the International Cancer Genome Consortium.

Next-generation DNA and RNA sequencing requires intact nucleic acids from high-quality human tissue samples to better elucidate the molecular basis of cancer. We have developed a prostate biobanking protocol to acquire suitable samples for sequencing without compromising the accuracy of clinical diagnosis. To assess the clinical implications of implementing this protocol, we evaluated 105 consecutive radical prostatectomy specimens from November 2008 to February 2009. Alternating levels of prostate samples were submitted to Surgical Pathology as formalin-fixed, paraffin-embedded blocks and to the institutional biobank as frozen blocks. Differences in reported pathologic characteristics between clinical and procured specimens were compared. Clinical staging and grading were not affected by the biobank protocol. Tumor foci on frozen hematoxylin and eosin slides were identified and high-density tumor foci were scored and processed for DNA and RNA extractions for sequencing. Both DNA and RNA were extracted from 22 cases of 44 with high-density tumor foci. Eighty-two percent (18/22) of the samples passed rigorous quality control steps for DNA and RNA sequencing. To date, DNA extracted from 7 cases has undergone whole-genome sequencing, and RNA from 18 cases has been RNA sequenced. This protocol provides prostate tissue for high-throughput biomedical research and confirms the feasibility of actively integrating prostate cancer into The Cancer Genome Atlas Program, a member of the International Cancer Genome Consortium.

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期刊介绍: Diagnostic Molecular Pathology focuses on providing clinical and academic pathologists with coverage of the latest molecular technologies, timely reviews of established techniques, and papers on the applications of these methods to all aspects of surgical pathology and laboratory medicine. It publishes original, peer-reviewed contributions on molecular probes for diagnosis, such as tumor suppressor genes, oncogenes, the polymerase chain reaction (PCR), and in situ hybridization. Articles demonstrate how these highly sensitive techniques can be applied for more accurate diagnosis.
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