Mutations in BRCA1 from fixed, paraffin-embedded tissue can be artifacts of preservation
Mutations in BRCA1 from fixed, paraffin-embedded tissue can be artifacts of preservation
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DOI：
10.1016/s0165-4608(98)00079-x
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发表时间：
1998-11-01
影响因子：
--
通讯作者：
Piver, MS
中科院分区：
文献类型：
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作者：
Wong, C;DiCioccio, RA;Piver, MS
从石蜡包埋的组织中分离的DNA已用于分析疾病状态中的DNA改变。使用存档组织可以加快收集大量罕见疾病亚型的标本，这些标本需要数年的时间才能积累。因此，检索了70例在40岁之前或40岁时诊断的卵巢癌病例的存档组织，用于BRCA1突变分析。从40岁之前或40岁时诊断的70例卵巢癌患者的石蜡包埋组织中分离DNA。采用单链构象多态性分析和DNA测序方法对BRCA1基因突变进行分析。在70例卵巢癌病例中，有34例发现了58个BRCA1突变。22例各有一个突变，12例有多个突变。2例组织学正常组织中发现的多个突变在匹配的肿瘤组织中不存在。在另一个案例中，来自两个不同正常组织块的DNA含有不同的突变。这些观察结果是异常的，表明在固定组织中检测到的突变可能是组织保存的伪影，而不存在于原始未固定组织中。为了验证这一建议，从2名在固定的正常组织中发现突变的患者中获得血液。来自未固定淋巴细胞的DNA不包含在固定的正常组织中发现的突变。因此，在固定的石蜡包埋组织中发现的突变可能是组织保存的伪影。在缺乏来自未固定组织的确证数据的情况下，来自这些组织的DNA序列数据的可靠性必须受到质疑。这严重限制了使用固定的组织作为DNA来源，用于回顾性研究和在患病成员不存活的家庭中进行临床基因检测。(C)Elsevier Science Inc.，1998.
DNA isolated from paraffin-embedded tissues has been used for analysis of DNA alterations in disease states. Use of archival tissue can expedite the gathering of large numbers of specimens from rare disease subtypes that would take years to accumulate prospectively. Therefore, archival tissues from 70 ovarian cancer cases diagnosed before or at age 40 were retrieved for analysis of BRCA1 mutations. DNA was isolated from paraffin-embedded tissue of 70 ovarian cancer cases diagnosed before or at age 40. BRCA1 mutation analysis was conducted by single-strand conformation polymorphism analysis and DNA sequencing. Fifty-eight BRCA1 mutations were found in 34 of the 70 ovarian cancer cases. Twenty-two cases had one mutation each and 12 cases had multiple mutations. Multiple mutations found in histologically normal tissue of 2 cases were not present in matched tumor tissue. For another case, DNA from two separate blocks of normal tissue contained different mutations. These observations were anomalous and suggested that mutations detected in fixed tissues may be artifacts of tissue preservation and not present in the original unfixed tissues. To test this suggestion, blood was obtained from 2 patients for whom mutations were found in fixed, normal tissue. DNA from their unfixed lymphocytes did not contain the mutations found in fixed normal tissue. Thus, mutations found in fixed, paraffin-embedded tissues can be artifacts of tissue preservation. The reliability of DNA sequence data derived from such tissues must be questioned in the absence of corroborating data from unfixed tissues. This severely limits the use of fixed tissues as a source of DNA for retrospective research and for clinical genetic testing in families for which a disease-affected member is not alive. (C) Elsevier Science Inc., 1998.