Allred Score Research Articles

Relationship between K-Ras mutational status and EGFR expression evaluated using Allred score in primary and metastatic colorectal cancer.

3568 Background: EGFR plays an important role in colorectal cancer (CRC) tumor progression and represents a natural target for molecular anticancer drugs. The monoclonal antibodies (MAbs) cetuximab and panitumumab, capable to inactivate the EGFR pathway, have entered in clinical practice for the treatment of metastatic CRC. While EGFR expression, detected by immunohistochemistry (IHC), is unable to predict response to molecular therapy, recent studies demonstrated that K-Ras status may be predictive of response to the MAbs. Methods: In this study we aimed to: 1. determine whether a correlation between K-Ras mutations and EGFR overexpression may occur, 2. verify whether K-Ras status may change during tumor progression. To this end in 414 CRC patients we evaluated K-Ras mutations using a direct cycle sequencing and EGFR expression using the Allred score which is a composite of the percentage of stained cells and the intensity of their staining. In 282 (68%) cases the analyses were performed in the primary tumor, in 67 (15%) in metastatic lesions and in 70 (17%) both in primary and in metastatic tumors. Results: we evidenced that 154 CRCs (37 %) were K-Ras mutated and 342 (82%) EGFR positive. In the 102 CRCs with EGFR negative or with a low Allred score (2-3) we found 65% of cases K-Ras mutated being mutated only 24% of the 73 CRCs with a high EGFR Allred score (7-8) (p = 0.0001). Moreover, we observed that 4 of 62 (6%) primary CRC and paired metastases showed changes in K-Ras mutational status. These cases did not vary in EGFR expression and did not show any differences concerning Loss of Heterozygosity (LOH) of CA Simple Sequence Repeat 1 (CASSRI) of intron 1 of EGFR. Conclusions: our data showed that CRC with K- Ras wild type consistently presents a high EGFR Allred score. Moreover, although K-Ras status varies between during progression in a small percentage of cases, a different response to monoclonal antibodies treatment may occur in these patients. Therefore, the characterization of the molecular profile in metastatic CRC has become increasingly important for targeted therapy selection. No significant financial relationships to disclose.

Osteopontin expression is a valuable marker for prediction of short-term recurrence in WHO grade I benign meningiomas

Prediction of recurrence remains a challenge in histopathological benign/grade I tumors. Osteopontin (OPN) plays important roles in tumorigenesis, invasion, and metastasis of several human cancers. In this study, we investigated OPN protein expression by evaluating the differences between recurrent and non-recurrent histologically benign meningiomas. Thirty-two patients were enrolled, and 23 benign non-recurrent meningiomas and 9 benign recurrent meningiomas were followed for a mean of 34months after complete surgical resection (Simpson grades I and II). Cytoplasmic OPN staining was evaluated by means of immunohistochemistry (IHC) score and by use of the Allred-8-unit system. We examined clinical biological data, their relationship with tumor recurrence, and the expression of OPN. Our results showed that meningioma recurrence correlated significantly with OPN IHC score (P=0.001). An OPN Allred score between 0 and 3 was associated with a recurrence-free time of more than 25months. In comparison, an OPN Allred score from 4 to 8 was indicative of a shorter average recurrence-free time. We concluded that OPN IHC score may play a role in prediction of the recurrence of the grade I meningiomas. Moreover, determination of the OPN Allred score is a reliable, quantitative tool for predicting recurrence-free time in benign meningioma patients.

Possible Difference in Frequencies of Genetic Polymorphisms of Estrogen Receptor , Estrogen Metabolism and P53 Genes Between Estrogen Receptor-positive and -negative Breast Cancers

Genetic polymorphisms associated with breast cancer risk are likely to differ among ethnic and molecular subtypes. The ability to identify genetic polymorphisms affecting the risk of estrogen receptor (ER)-positive breast cancer may lead to the more efficient selection of candidates for chemoprevention with endocrine agents. We focused on identifying common genotypes for ER-positive breast cancer in premenopausal Japanese women. We compared genetic polymorphisms of ERalpha, estrogen metabolism genes (CYP17A1, CYP19A1, HSD17B1 COASY, CYP1B1 and COMT), and p53 between ER-positive and -negative female Japanese breast cancer patients, and analyzed whether these polymorphisms affected the frequency of ER-positive breast cancer. Carriers of the G allele of ERalpha (rs6905370) were more frequent in ER-positive breast cancer than in ER-negative breast cancer especially in those under 50-year old. Pairwise analysis showed that combinations of the ERalpha G allele with the homozygous Trp genotype of CYP19A1 codon 39 (rs2236722), the methionine (Met) allele of COMT codon 158 (rs4680) or Pro allele of p53 codon 72 (rs1042522) were more frequent in ER-positive than ER-negative breast cancer, especially in patients less than 50-year old. The frequencies of these combinations were even higher in patients with strongly ER-positive tumors (Allred's scores of 7 or 8). Our study demonstrated genetic polymorphisms of ERalpha, CYP19A1, COMT and p53 genes frequently occur in ER-positive breast cancer in premenopausal Japanese women.

Problems and Solutions in the Evaluation of Hormone Receptors in Breast Cancer

Problems and Solutions in the Evaluation of Hormone Receptors in Breast Cancer

A randomised study of the effects of letrozole and anastrozole on oestrogen receptor positive breast cancers in postmenopausal women

Changes in proliferation as measured by Ki67 occur within 14 days of starting treatment with an aromatase inhibitor and these changes have been shown to be predictors of long term outcome. This study aimed to compare changes in proliferation following 14 days of treatment with anastrozole and letrozole. Two hundred and six women with 209 estrogen receptor (ER) positive operable breast cancers (three bilateral) were randomly allocated to receive either 14 days treatment with 2.5 mg of letrozole or 1 mg of anastrozole prior to surgery. Changes in expression of estrogen (ER) and progesterone receptors (PgR) as assessed by ALLRED scores and proliferation as assessed by Ki67 were analysed. The HER2 status of each tumour was also assessed using a combination of the Hercept test and FISH. Both letrozole and anastrozole reduced ER expression (ALLRED score) by a mean of 0.32 (0.20-0.44), P<0.001 and PgR fell by a mean of 2.54 (2.20-2.89) P<0.0001. Letrozole reduced proliferation from a geometric mean of 6.37% to 0.81%, P<0.0001 and anastrozole reduced proliferation from 5.81% to 0.77%, P<0.0001. There was no differences between drugs in the fall in ER, PgR or proliferation. Both letrozole and anastrozole produced significant falls in proliferation in both HER2 positive and HER2 negative cancers, all P<0.001. 14 days of both letrozole and anastrozole reduces proliferation, ER and PgR expression. No significant difference between these two drugs was identified.

Development of Standard Estrogen and Progesterone Receptor Immunohistochemical Assays for Selection of Patients for Antihormonal Therapy

Estrogen receptor (ER) and progesterone receptor (PR) status in breast carcinomas are considered validated predictive factors for selecting patients for antihormonal therapy. Published surveys have shown a significant rate of disagreement and lack of reproducibility of immunohistochemistry (IHC) results from laboratories around the world. To address these limitations IHC assays for ER and PR were developed using characterized reagents, after careful calibration of the sensitivity and specificity to match established assays previously validated in large clinical studies. The ER assay uses a cocktail of 2 mouse monoclonal antibodies (1D5 and ER-2-123) and the PR assay uses 1 mouse monoclonal antibody (PgR 1294); both are followed by a polymer-peroxidase-based detection system. All antibodies were tested for specificity by epitope mapping. The sensitivity of the new assays was calibrated to be equivalent to previously validated IHC assays followed by a comparison with the validated assays in a concordance study involving over 200 specimens. All slides were scored with the "Allred Score," also used for scoring of the original validated assays. The overall concordance between the new and the established IHC assays was nearly perfect (99%). The concordance study demonstrated greater than 98% positive agreement and 100% negative agreement of the new IHC assays with the previously validated IHC assays. This equivalence establishes the clinical validation of the assays and, as they are based on newer generation reagents and are produced and tested under stringent quality control conditions to ensure their consistency, they add additional advantages to the user and patients.

Receptor for advanced glycation end products (RAGE) is important in the prediction of recurrence in human oral squamous cell carcinoma

Receptor for advanced glycation end products (RAGE) has recently been recognized as a cancer-associated protein responsible for cancer progression and metastasis in gastrointestinal cancers. The aim was to examine the role of RAGE in oral squamous cell carcinoma (OSCC). RAGE expression was examined by immunohistochemistry in 74 OSCC patients and evaluated with a grading based on Allred's score. RAGE expression was compared with clinicopathological parameters including clinical stage, invasive depth, nodal metastasis, disease recurrence and disease-free survival. High-grade expression of RAGE (RAGE-H) was observed in 30 (40.5%) of 74 OSCCs. RAGE-H was associated with depth of invasion (P < 0.0001) and local recurrence (P < 0.0001), but not with histological differentiation, clinical stage or nodal metastasis. Disease-free survival in patients with RAGE-H was significantly worse than in those with low-level RAGE expression. Multivariate analysis showed RAGE-H to be an independent prognostic factor for disease-free survival in OSCC patients (P = 0.0022). RAGE is a relevant factor in predicting disease recurrence and patients' prognosis in OSCC.

Concordance of local and central laboratory hormone and HER2 receptor status in ECOG 2197

21022 Background: Central and local laboratory concordance for hormone and HER2 receptor measurement is of national interest. This study compares ER/PR/HER2 by local laboratories using immunohistochemistry (IHC) and central laboratories (IHC &amp; quantitative RT-PCR). Methods: Of 2952 patients in E2197, a case-cohort sample of 776 patients who either did (N=179) or did not recur was studied. Central IHC for ER/PR/HER2 was performed using single 0.6 mm microarrays; Allred score (AS) was used for ER/PR (AS&gt;2 = positive). Positive HER2 was 3+ staining in &gt;10% cells for Central IHC and 2+ or 3+ for Local IHC. RT-PCR analysis by Oncotype DX™ for ER/PR/HER2 was performed using pre-defined cutoffs of 6.5, 5.5 and 11.5 units, respectively. Hormone receptor (HR) pos was defined as ER &amp;/or PR pos. Results: Results from Local IHC (ER/PR in 776 &amp; HER2 in 517 pts) were compared with Central IHC (760 pts) and RT-PCR results (776 pts). The discordance between HR positivity by Local IHC and RT-PCR was very low. However, 12% of HR neg pts by Local IHC (38/321) &amp; Central IHC (39/326) were HR pos by RT-PCR. The relationship between ER and recurrence as a function of AS was examined. Patients with AS of 3–4 were found to be closer to the AS=2 group than to the AS&gt;4 group Patients with AS of 3–4 were found to be closer to the AS ÿ 2 group than to the AS &gt; 4 group (Est.HR for ER 0.97 for AS 3–4 vs. 0–2 and 0.46 for AS 5–8 vs. 0–2, and for PR were 0.84 for AS 3–4 vs. 0–2 and 0.41 for AS 5–8 vs. 0–2). Conclusions: There is a high degree of overall concordance among Local IHC, Central IHC, and Central RT-PCR for ER and PR. The degree of concordance is even greater for HR compared to ER or PR alone. Although the concordance with local labs for HER2 testing was poor, the concordance between Central IHC and RT-PCR was very high. The relatively high incidence (12%) of IHC HR neg pts who are HR pos by RT- PCR is notable. [Table: see text] [Table: see text]

Immunohistochemical evaluation of hormone receptor status for predicting response to endocrine therapy in metastatic breast cancer

The importance of establishing hormone receptor status of tumors for the treatment of women with hormone receptor-positive breast cancer has been emphasized, however, there is no general agreement as to how immunohistochemical assays should be evaluated. It is critical to evaluate hormone receptor status when considering response to endocrine therapy. Estrogen receptor (ER) and progesterone receptor (PgR) expression was examined by immunohistochemistry using Allred's score for primary breast tumors from 75 metastatic breast cancer patients who received first-line treatment with endocrine therapy (56 patients received tamoxifen, 11 patients received aromatase inhibitors, and 8 patients received LH-RH agonist or other endocrine reagents) on relapse. Correlation between hormone receptor status and response to endocrine therapy as well as post-relapse survival was analyzed. The most significant correlation between positive ER expression and response to any endocrine therapy (p = 0.011) or tamoxifen only (p = 0.030) occurred when the cutoff score was set at 10%. When the evaluation was based on Allred's score (TS), a cutoff point of TS>or=4 showed a more significant association between positive ER expression and response to all kinds of endocrine therapy (p = 0.020) or tamoxifen only (p = 0.047). When evaluated at a cutoff point of 1% positive cells, there were fifteen patients with both ER- and PgR-negative tumors, and three patients (20.0%) responded to the therapy. Patients with 1% or more ER or PgR positive cells had better survival after relapse (p = 0.0005 and p = 0.0008, respectively). The proportion score alone might be enough to predict hormone responsiveness and post-relapse survival in metastatic breast cancer. The cutoff might be set low, for example 1%, especially for metastatic disease.

Immunohistochemical assessment for estrogen receptor and progesterone receptor status in breast cancer: Analysis for a cut-off point as the predictor for endocrine therapy

An immunohistochemical (IHC) method is commonly used for determining estrogen receptor (ER) and progesterone receptor (PR) status in breast cancer. However, the proper cut-off points of IHC have not been established. Cut-off points for ER and PR status as predictive factors for endocrine therapy are needed. A total of 249 cases of female breast cancer were enrolled. ER and PR status by IHC were analyzed using the proportion of stained cells and staining intensity by Allred's score. Proportion score (PS) and intensity score (IS) were related to enzyme immunoassay (EIA) titers, for both in ER and PR (p < 0.0001, all). PS correlated with IS in both ER and PR (R = 0.47 and 0.41, respectively). ER status by IHC was related to tumor size and lymph node status, while PR was related to tumor size and menopausal status. In 152 patients who received endocrine therapy with a median follow-up term of 38 months, differences in disease-free survival were most significant using a cut-off point of PS 3 which indicated more than 10 % of cells stained positively for both ER and PR (p = 0.0007 and 0.0087, respectively). In addition, combination analysis of ER and PR using this cut-off point revealed a notable prognostic difference. A 10 % staining proportion may be an acceptable cut-off point for both ER and PR status by IHC, in terms of predicting response to endocrine therapy in breast cancer.