Assessment of Tissue Estrogen and Progesterone Receptor Levels: A Survey of Current Practice, Techniques, and Quantitation Methods

Abstract

The assessment of steroid hormone receptors in resected breast carcinoma tissue is currently the standard of practice. The traditional method for assessment of receptor status is the ligand binding assay. More recently, immunohistochemistry (IHC) has become a popular method for such testing. Despite the widespread use of IHC and the availability of many antibodies, standardization of quantitative IHC for assessment of estrogen and progesterone receptors has not been achieved. While the College of American Pathologists (CAP) offers a Quality Assurance (QA) program for IHC quantitation of estrogen receptor (ER) and progesterone receptor (PgR), no universal standard is currently recognized in assessment of ER and PgR by IHC. We surveyed 300 laboratories within the United States for their current practices regarding the assessment of ER and PgR status in breast cancer tissue specimens. Eighty usable responses were received. Forty-nine (61%) laboratories performed the assay in-house, while the remainder sent the material out for assay. All responding laboratories performing their steroid receptor analysis in-house used the IHC technique. Forty-three (80%) laboratories answering the question on material accepted for analysis performed the assay only on paraffin-embedded material, three (6%) used either paraffin block or frozen material, and two (4%) used only frozen material. Eighty-eight percent of laboratories performing steroid receptor analysis in-house used a manual quantitation technique. Four (8%) used computer-assisted image analysis, and a single laboratory used laser scanning cytometry. Eight different antibodies were used among the 44 laboratories documenting the antibody supplier, and for any given commercially prepared antibody a wide variety of dilutions were used, with the exception of the standard solution used with the Ventana antibody. Of the laboratories using manual estimation techniques, 61% simply estimated the percentage of positive cells, 29% evaluated both the intensity of staining and percentage of nuclei staining, 6% used formal H-score analysis, 2% evaluated only intensity of nuclear staining, and 2% mainly counted the percentage of nuclei staining for ER but used a formal H score in the assessment of PgR. Cutoff points for the separation of positive and negative results varied widely, with some laboratories assessing any demonstrable positivity as a positive result, while others required as many as 19% of the nuclei to stain before a specimen was declared positive. Standardization techniques differed considerably among laboratories. Eighty-six percent used the CAP program for QA. While all laboratories utilized some form of intralaboratory control for assessment of ER and PgR, the nature of that control varied from laboratory to laboratory. Our survey indicates that a majority of laboratories perform their steroid hormone receptor analysis in-house using IHC. There is considerable variability in the antibodies utilized, the dilutions applied, and the quantitation method and level of express...

Diagnostic value of diffusion‑weighted imaging‑derived apparent diffusion coefficient and its association with histological prognostic factors in breast cancer

“…The ADC mean for malignant and benign tumors in the present study were 1.0±0.2 and 1.5±0.2, respectively. This was, to a certain degree, consistent with the results of previous studies on breast lesions (9,17,20,21). Compared with previous studies, the present study included a large population and yielded more comprehensive results.…”

Diagnostic value of diffusion‑weighted imaging‑derived apparent diffusion coefficient and its association with histological prognostic factors in breast cancer

“…The ADC mean for malignant and benign tumors in the present study were 1.0±0.2 and 1.5±0.2, respectively. This was, to a certain degree, consistent with the results of previous studies on breast lesions (9,17,20,21). Compared with previous studies, the present study included a large population and yielded more comprehensive results.…”

Assessment of hormone receptor status in breast cancer

“…The present retrospective study examined a unique group of patients who underwent postoperative tamoxifen monotherapy. The best cut‐off point thus could not be established based solely on the results of the present study, but it is interesting that significance was identified for a cut‐off of 10%, which is used in many hospitals 5 …”

Is Oncotype DX testing informative for breast cancers with low ER expression? A retrospective review from a biomarker testing referral center

“…False positive ER-low results could lead to suboptimal adjuvant treatment and it is therefore important to consider preanalytical factors, such as cold ischemic time and specimen fixation protocols, either of which could contribute to falsely low ER staining intensity. There are also well-recognized analytic factors influencing the consistency of ER intensity reporting, most of which arise from different laboratories using different and closed-system protocols for tissue processing and staining [ 8 , 22 , 23 ]. A lack of standardized ER intensity thresholds was one of the principal contributing factors in at least one highly publicized instance involving significant under-reporting of ER-positive cases [ 24 ].…”

Diagnostic value of diffusion‑weighted imaging‑derived apparent diffusion coefficient and its association with histological prognostic factors in breast cancer

“…The ADC mean for malignant and benign tumors in the present study were 1.0±0.2 and 1.5±0.2, respectively. This was, to a certain degree, consistent with the results of previous studies on breast lesions (9,17,20,21). Compared with previous studies, the present study included a large population and yielded more comprehensive results.…”

Assessment of hormone receptor status in breast cancer

“…The present retrospective study examined a unique group of patients who underwent postoperative tamoxifen monotherapy. The best cut‐off point thus could not be established based solely on the results of the present study, but it is interesting that significance was identified for a cut‐off of 10%, which is used in many hospitals 5 …”

Is Oncotype DX testing informative for breast cancers with low ER expression? A retrospective review from a biomarker testing referral center

“…False positive ER-low results could lead to suboptimal adjuvant treatment and it is therefore important to consider preanalytical factors, such as cold ischemic time and specimen fixation protocols, either of which could contribute to falsely low ER staining intensity. There are also well-recognized analytic factors influencing the consistency of ER intensity reporting, most of which arise from different laboratories using different and closed-system protocols for tissue processing and staining [ 8 , 22 , 23 ]. A lack of standardized ER intensity thresholds was one of the principal contributing factors in at least one highly publicized instance involving significant under-reporting of ER-positive cases [ 24 ].…”

Diagnostic value of diffusion‑weighted imaging‑derived apparent diffusion coefficient and its association with histological prognostic factors in breast cancer

“…The ADC mean for malignant and benign tumors in the present study were 1.0±0.2 and 1.5±0.2, respectively. This was, to a certain degree, consistent with the results of previous studies on breast lesions (9,17,20,21). Compared with previous studies, the present study included a large population and yielded more comprehensive results.…”

Assessment of hormone receptor status in breast cancer

“…The present retrospective study examined a unique group of patients who underwent postoperative tamoxifen monotherapy. The best cut‐off point thus could not be established based solely on the results of the present study, but it is interesting that significance was identified for a cut‐off of 10%, which is used in many hospitals 5 …”

Is Oncotype DX testing informative for breast cancers with low ER expression? A retrospective review from a biomarker testing referral center

“…False positive ER-low results could lead to suboptimal adjuvant treatment and it is therefore important to consider preanalytical factors, such as cold ischemic time and specimen fixation protocols, either of which could contribute to falsely low ER staining intensity. There are also well-recognized analytic factors influencing the consistency of ER intensity reporting, most of which arise from different laboratories using different and closed-system protocols for tissue processing and staining [ 8 , 22 , 23 ]. A lack of standardized ER intensity thresholds was one of the principal contributing factors in at least one highly publicized instance involving significant under-reporting of ER-positive cases [ 24 ].…”