Esgueva Raquel, Park Kyung, Kim Robert, Kitabayashi Naoki, Barbieri Christopher E, Dorsey Philip J, Abraham Cyril, Banerjee Samprit, Leung Robert A, Tewari Ashutosh K, Terry Stéphane, Shevchuk Maria M, Rickman David S, Rubin Mark A
Departments of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY 10065, USA.
Diagn Mol Pathol. 2012 Jun;21(2):61-8. doi: 10.1097/PDM.0b013e31823b6da6.
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.
新一代DNA和RNA测序需要来自高质量人体组织样本的完整核酸，以便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样本库方案，以获取适合测序的样本，同时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，我们对2008年11月至2009年2月期间连续的105例根治性前列腺切除术标本进行了评估。前列腺样本的交替水平分别作为福尔马林固定、石蜡包埋块提交给外科病理学，并作为冷冻块提交给机构生物样本库。比较了临床标本和获取标本之间报告的病理特征差异。临床分期和分级不受生物样本库方案的影响。在冷冻苏木精和伊红切片上识别肿瘤灶，对高密度肿瘤灶进行评分，并处理用于DNA和RNA提取以进行测序。在44例有高密度肿瘤灶的病例中，从22例中提取了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，从7例中提取的DNA已进行全基因组测序，18例的RNA已进行RNA测序。该方案为高通量生物医学研究提供了前列腺组织，并证实了将前列腺癌积极纳入国际癌症基因组联盟成员之一的癌症基因组图谱计划的可行性。
