PAM50 breast cancer subtyping by RT-qPCR and concordance with standard clinical molecular markers.
PAM50 breast cancer subtyping by RT-qPCR and concordance with standard clinical molecular markers.
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DOI：
10.1186/1755-8794-5-44
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发表时间：
2012-10-04
影响因子：
2.7
通讯作者：
Martín M
中科院分区：
文献类型：
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作者：
Bastien RR;Rodríguez-Lescure Á;Ebbert MT;Prat A;Munárriz B;Rowe L;Miller P;Ruiz-Borrego M;Anderson D;Lyons B;Álvarez I;Dowell T;Wall D;Seguí MÁ;Barley L;Boucher KM;Alba E;Pappas L;Davis CA;Aranda I;Fauron C;Stijleman IJ;Palacios J;Antón A;Carrasco E;Caballero R;Ellis MJ;Nielsen TO;Perou CM;Astill M;Bernard PS;Martín M
研究中使用了许多方法来鉴定乳腺癌的“内在”亚型，通常称为管腔A型、管腔B型、HER 2富集型（HER 2-E）和基底样型。PAM 50基因集通常用于基于基因表达的亚型分型;然而，使用免疫组织化学（IHC）标记物的替代亚型分型仍然在临床上广泛使用。这些方法之间的差异可能导致不同的治疗决定。我们使用PAM 50 RT-qPCR检测对来自GEICAM/9906 III期临床试验的814个肿瘤进行表达谱分析，该试验招募了患有局部晚期原发性浸润性乳腺癌的女性。通过IHC在单个位点对所有样本的雌激素受体（ER）、孕酮受体（PR）和Her 2/neu（HER 2）蛋白表达进行评分。通过显色原位杂交（CISH）证实了可疑的HER 2病例。将通过IHC/CISH的单基因评分与RT-qPCR连续基因表达值和通过PAM 50的“内在”亚型分配进行比较。使用来自PAM 50亚型分配的连续RT-qPCR数据的四分位数临界点，选择ESR 1、PGR、ERBB 2和增殖的高、中和低表达。ESR 1、PGR和ERBB 2基因表达与确定的二进制IHC临界点（曲线下面积（AUC）≥ 0.9）高度一致。通过IHC检测的雌激素受体阳性与Luminal（A和B）亚型（92%）密切相关，但只有75%的ER阴性肿瘤分为HER 2-E和基底样亚型。管腔A型肿瘤比管腔B型肿瘤更常表达PR（94% vs 74%），管腔A型肿瘤不太可能具有高增殖（11% vs 77%）。77%（30/39）的ER-/HER 2+肿瘤通过IHC被分类为HER 2-E亚型。三阴性肿瘤主要包括基底细胞样（57%）和HER 2-E（30%）亚型。如多变量分析所示，ESR 1、PGR和ERBB 2的单基因评分比相应的IHC标志物更具预后性。乳腺癌（ER、PR和HER 2）的标准免疫组化组不能充分识别PAM 50基因表达亚型。尽管通过蛋白质免疫组织化学和基因表达的生物标志物评分之间存在高度一致性，但ESR 1和ERBB 2状态的基因表达测定更具预后性。
Many methodologies have been used in research to identify the “intrinsic” subtypes of breast cancer commonly known as Luminal A, Luminal B, HER2-Enriched (HER2-E) and Basal-like. The PAM50 gene set is often used for gene expression-based subtyping; however, surrogate subtyping using panels of immunohistochemical (IHC) markers are still widely used clinically. Discrepancies between these methods may lead to different treatment decisions. We used the PAM50 RT-qPCR assay to expression profile 814 tumors from the GEICAM/9906 phase III clinical trial that enrolled women with locally advanced primary invasive breast cancer. All samples were scored at a single site by IHC for estrogen receptor (ER), progesterone receptor (PR), and Her2/neu (HER2) protein expression. Equivocal HER2 cases were confirmed by chromogenic in situ hybridization (CISH). Single gene scores by IHC/CISH were compared with RT-qPCR continuous gene expression values and “intrinsic” subtype assignment by the PAM50. High, medium, and low expression for ESR1, PGR, ERBB2, and proliferation were selected using quartile cut-points from the continuous RT-qPCR data across the PAM50 subtype assignments. ESR1, PGR, and ERBB2 gene expression had high agreement with established binary IHC cut-points (area under the curve (AUC) ≥ 0.9). Estrogen receptor positivity by IHC was strongly associated with Luminal (A and B) subtypes (92%), but only 75% of ER negative tumors were classified into the HER2-E and Basal-like subtypes. Luminal A tumors more frequently expressed PR than Luminal B (94% vs 74%) and Luminal A tumors were less likely to have high proliferation (11% vs 77%). Seventy-seven percent (30/39) of ER-/HER2+ tumors by IHC were classified as the HER2-E subtype. Triple negative tumors were mainly comprised of Basal-like (57%) and HER2-E (30%) subtypes. Single gene scoring for ESR1, PGR, and ERBB2 was more prognostic than the corresponding IHC markers as shown in a multivariate analysis. The standard immunohistochemical panel for breast cancer (ER, PR, and HER2) does not adequately identify the PAM50 gene expression subtypes. Although there is high agreement between biomarker scoring by protein immunohistochemistry and gene expression, the gene expression determinations for ESR1 and ERBB2 status was more prognostic.