HER-2/neu amplification testing in breast cancer by multiplex ligation-dependent probe amplification in comparison with immunohistochemistry and in situ hybridization
HER-2/neu amplification testing in breast cancer by multiplex ligation-dependent probe amplification in comparison with immunohistochemistry and in situ hybridization
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DOI：
10.3233/clo-2009-0461
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发表时间：
2009-01-01
影响因子：
6.6
通讯作者：
van Diest, Paul J.
中科院分区：
文献类型：
--
作者：
Moelans, Cathy B.;de Weger, Roel A.;van Diest, Paul J.
背景资料：评估HER-2/neu在浸润性乳腺癌中的状态对于确定曲妥珠单抗和紫杉烷类化疗的合格性至关重要。免疫组织化学（IHC）评估蛋白质过度表达，二线基因扩增测试是需要与可疑的蛋白质表达的情况下。本研究旨在验证一种新的基于PCR的检测，称为多重连接依赖性探针扩增（MLPA），作为一种简单快速的方法来评估浸润性乳腺癌HER-2/neu基因扩增状态。方法：将MPLA结果与荧光原位杂交（FISH）和显色原位杂交（CISH）评估的基因扩增状态进行比较，结果：518例乳腺癌组织中HER-2/neu蛋白表达阳性率为10%，MLPA检测HER-2/neu基因扩增阳性率为11%。FISH与CISH、MLPA与IHC、MLPA与CISH之间的一致性较高。MLPA在7/36例（19%）可疑IHC 2+病例中显示扩增。然而，在IHC 0/1+病例中，6/434例（1.4%）也通过MLPA扩增，并且所有这些病例均通过FISH/CISH证实扩增。结论：MLPA检测乳腺癌HER-2/neu扩增是一种快速、准确、廉价的方法，可作为FISH和CISH的可靠替代方法。
Background: Assessment of HER-2/neu status in invasive breast cancer is crucial to establish eligibility for trastuzumab and taxane based chemotherapy. Next to immunohistochemistry (IHC) to evaluate protein overexpression, a second line gene amplification test is required for cases with equivocal protein expression. This study aimed to validate a new PCR based test, called Multiplex Ligation-dependent Probe Amplification (MLPA), as a simple and quick method to assess HER-2/neu gene amplification status in invasive breast cancer.Methods: MPLA results were compared with gene amplification status assessed by fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) as gold standard, and with protein overexpression by IHC in 518 breast carcinoma patients.Results: About 10% of cases overexpressed HER-2/neu at the protein level (IHC), and 11% of cases showed gene-amplification by MLPA. A high concordance was found between FISH and CISH, MLPA and IHC, and MLPA and CISH. MLPA showed amplification in 7/36 (19%) of the equivocal IHC 2+ cases. However, of the IHC 0/1+ cases, 6/434 (1.4%) were also amplified by MLPA, and amplification was confirmed in all of these cases by FISH/CISH. On the other hand, one of the 48 (2%) IHC 3+ cases was normal by MLPA and lack of amplification was confirmed by FISH/CISH.Conclusion: MLPA is a fast, accurate and cheap method to detect breast cancer HER-2/neu amplification in small quantities of DNA extracted from paraffin blocks, and thereby a reliable alternative to FISH and CISH.