Allred Scoring System Research Articles

The Significance of TGF-β Expression in Predicting Lymphovascular Invasion and Lymph Node Metastasis in Colorectal Cancer

Background: Colorectal cancer (CRC) is a major health burden globally. The prognosis of CRC is strongly influenced by the presence of lymphovascular invasion (LVI) and lymph node (LN) metastasis. Transforming growth factor-beta (TGF-β) is a cytokine with a complex role in CRC progression. This study aimed to evaluate the significance of TGF-β expression in predicting LVI and LN metastasis in CRC. Methods: This cross-sectional study involved 50 patients diagnosed with CRC. The expression of TGF-β was assessed using immunohistochemical staining and the Allred scoring system. The relationship between TGF-β expression and the presence of LVI and LN metastasis was analyzed using the Chi-square test. Results: High TGF-β expression was significantly associated with both LVI (p = 0.011) and LN metastasis (p = 0.012) in CRC. Patients with high TGF-β expression had a higher risk of LVI and LN metastasis compared to those with low TGF-β expression. Conclusion: TGF-β expression is a significant predictor of LVI and LN metastasis in CRC. This finding has potential implications for risk stratification and treatment decisions in CRC patients.

Optimizing assessment of CD30 expression in Hodgkin lymphoma by controlling for low expression.

Since the approval of brentuximab vedotin (BV), assessment of CD30 status by immunohistochemistry gained increasing importance in the clinical management of patients diagnosed with CD30-expressing lymphomas, including classical Hodgkin lymphoma (CHL). Paradoxically, patients with low or no CD30 expression respond to BV. This discrepancy may be due to lack of standardization in CD30 staining methods. In this study, we examined 29 cases of CHL and 4 cases of nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) for CD30 expression using a staining protocol that was designed to detect low CD30 expression levels, and an evaluation system similar to the Allred scoring system used for breast cancer evaluation. For CHL, 10% of cases had low scores and 3% were CD30 negative, with 3 cases in which the majority of tumor cells showed very weak staining. Unexpectedly, one of four cases of NLPHL was positive. We demonstrate intra-patient heterogeneity in CD30 expression levels and staining patterns in tumor cells. Three CHL cases with weak staining may have been missed without the use of control tissue for low expression. Thus, standardization of CD30 immunohistochemical staining with use of known low-expressing controls may aid in proper CD30 assessment and subsequent therapeutic stratification of patients.

BRAF-V600E Protein Expression in Canine Malignant Cutaneous Melanoma, in Accordance with the Introduction of Biomarkers in Comparative Oncology Studies

Background: Melanoma is the cause of death for 1.3% of all cancer patients in humans. The key role of the BRAF protein in the progression of human melanoma has been confirmed, and its prognostic significance has been revealed. Because canine cancer resembles human cancer in biological behavior and molecular abnormalities, BRAF protein may be expressed in canine melanoma, the same as human melanoma. Despite the investigation of the BRAF mutation in canine melanoma, the status of BRAF at the protein level in canine skin melanoma has not yet been examined.
 Methods: Thirty-two formalin-fixed, paraffin-embedded tissue samples of canine malignant cutaneous melanoma were randomly selected. After cutting into 3-μmthick sections, the samples were evaluated for BRAF protein expression by immunohistochemistry and using the anti-BRAF V600E (VE1) mouse monoclonal antibody
 Results: The BRAF status was assessed using the Allred scoring system. Among the 32 samples examined, 21 samples were negative, and 11 cases showed high BRAF protein expression.
 Conclusion: The detection of positive BRAF expression in 34.3% of canine cutaneous melanoma samples could be a step forward to improving treatment options, using dogs as an animal model in human melanoma clinical trials, and possibly identifying a new prognostic biomarker in canine melanoma.

Clinicopathologic characterization of hepatocellular adenomas in men: a multicenter experience

Hepatocellular adenomas (HCAs) are benign liver neoplasms which most commonly present in women in their reproductive age. In men they are rare and have a higher risk of malignant transformation to hepatocellular carcinoma (HCC), with current guidelines advocating their complete resection. Here we present our multicenter experience of HCA in men in the United States. A total of 27 HCA cases were included with a mean age of presentation of 37 years (9 - 69 years) and a mean size of 6.8 cm (0.9 - 18.5 cm). Based on the 2019 WHO classification, the most common subtype identified was inflammatory HCA (IHCA) (10 cases, 37.0%) followed by unclassified HCA (U-HCA) (7 cases, 25.9%), HNF1A-inactivated HCA (H-HCA) (6 cases, 22.2%), β-catenin activated IHCA (b-IHCA) (3 cases, 11.1%), and β-catenin activated HCA (b-HCA) (1 case, 3.7%). Six additional cases diagnosed as hepatocellular neoplasm of uncertain malignant potential (HUMP) were also included in the study. These cases presented in a mean age of 46 years (17 - 64 years) and size of 10.8 cm (4.2 - 16.5 cm). We further evaluated the significance of androgen receptor (AR) expression by immunohistochemistry (IHC); of the 16 cases with materials available, 8 were considered positive using the Allred score system (2 I-HCA, 2 H-HCA, 1 U-HCA, and 3 HUMP). Of the total cases, 12 were diagnosed on biopsies, for which follow-up information is available for 7 (47.6 months, 8 - 160 months), and none of them show evidence of malignant transformation. Of the 21 resection cases, a concomitant well differentiated hepatocellular carcinoma (HCC) within the same lesion was identified in 5 cases (23.8%) which were diagnosed as HCA (n=4) or HUMP (n=1). Overall, 15% of cases in our entire cohort of HCA and HUMP showed concomitant HCC, while none of the 7 biopsy cases showed any malignant transformation on follow-up (22 - 160 months; mean 61.8).

Associations of CTCF and FOXA1 with androgen and IGF pathways in men with localized prostate cancer

AimsTo examine associations between the transcription factors CCCTC-binding factor (CTCF) and forkhead box protein A1 (FOXA1) and the androgen receptor (AR) and their association with components of the insulin-like growth factor (IGF)-pathway in a cohort of men with localized prostate cancer. MethodsUsing prostate tissue samples collected during the Prostate cancer: Evidence of Exercise and Nutrition Trial (PrEvENT) trial (N = 70 to 92, depending on section availability), we assessed the abundance of CTCF, FOXA1, AR, IGFIR, p-mTOR, PTEN and IGFBP-2 proteins using a modified version of the Allred scoring system. Validation studies were performed using large, publicly available datasets (TCGA) (N = 489). ResultsWe identified a strong correlation between CTCF and AR staining with benign prostate tissue. CTCF also strongly associated with the IGFIR, with PTEN and with phospho-mTOR. FOXA1 was also correlated with staining for the IGF-IR, with IGFBP-2 and with staining for activated phosphor-mTOR. The staining for the IGF-IR was strongly correlated with the AR. ConclusionOur findings emphasise the close and complex links between the endocrine controls, well known to play an important role in prostate cancer, and the transcription factors implicated by the recent genetic evidence.

Prognostic Value of CXCL12 in Non-Small Cell Lung Cancer Patients Undergoing Tumor Resection

Adjuvant chemotherapy is commonly indicated in lung cancer patients undergoing surgical therapy because tumor recurrence is frequent. A biomarker that can predict tumor recurrence in the postoperative period is currently unavailable. CXCR4 receptor and its ligand CXCL12 play important roles in metastasis. This study investigated the value of tumor CXCL12 expression to predict prognosis and indicate adjuvant chemotherapy in non-small cell lung cancer patients. This study enrolled 82 non-small cell lung cancer patients. The expression of CXCL12 was evaluated by immunohistochemistry. The degree of CXCL12 expression was assessed using the Allred score system. Among all subjects, the progression-free survival and overall survival were significantly prolonged in cancer patients with low tumor expression of CXCL12 compared to patients with high tumor expression. Multivariate analysis showed that the increased level of CXCL12 is a significant predictor of progression-free survival and overall survival in NSCLC patients. Among subjects with high tumor CXCL12 expression, progression-free survival and overall survival were significantly improved in patients treated with adjuvant chemotherapy compared to untreated patients. These results suggest the potential value of tumor CXCL12 expression as a marker to predict prognosis and to indicate adjuvant chemotherapy after surgical tumor resection in non-small cell lung cancer patients.

Estrogen Receptor Expression Pattern in Papillary Thyroid Cancers: A 23 Years Retrospective Multi-Centre Study in Jos Metropolis, North-Central, Nigeria

Abstract Introduction/Objective Papillary Thyroid cancers (PTC) are the commonest malignancy of this organ and seen more often in females. Estrogen Receptor (ER) is important in the pathogenesis of the disease and impacts on the prognosis. The aim is to establish ER status of PTC seen at the Jos University Teaching Hospital and other health institutions in Jos Metropolis. Methods/Case Report This was a hospital based, retrospective multicenter study. The biodata of histologically diagnosed PTC were obtained from surgical pathology cancer registries and archived histology request forms between 1st January 1998 and 31st December, 2020. Archival slides and fresh sections from tissue blocks were reviewed, classified and subjected to immunohistochemistry (polymer technique). They were scored (using Allred scoring system) and data analyzed using SPSS-20. Results (if a Case Study enter NA) A total of 53 (56%) cases of histologically diagnosed PTC out of 95 cases of thyroid cancers were included in the study. The mean, mode, median and age range were 44.7±15.4 years, 45 years, 43 years and 13-80 years respectively. Majority of cases were females (81.1%), this was replicated across all age groups. Out of the total number of PTC cases recorded, 36 (67.9%) were classical variant while the remaining 17 (32.1%) were follicular variants. Nine cases (17%) were positive for ER while 44 (83%) were negative. Out of the positive cases, 7 (77.8%) were females with 2 (22.2%) males. The total scores assigned to positive cases were 3 (66.7%) and 4 (33.3%) and were predominant in females in the 4th decade. Conclusion Majority of PTC were seen in females and were negative for ER. This suggest that very few of our patients are likely to benefit from selective estrogen receptor targeted therapy and therefore Surgery, Chemotherapy and Radioiodine therapy remain the main stay of treatment for most cases in our locality.

Utility of Secretagogin as a Novel Marker for the Diagnosis of Lung Neuroendocrine Carcinoma

BackgroundSmall cell lung cancers (SCLC) and large cell neuroendocrine carcinomas (LCNEC) are a subset of high‐grade lung carcinomas with characteristic morphology and molecular profile. Commonly used neuroendocrine markers include synaptophysin, chromogranin A, CD56, and insulinoma‐associated protein 1 (INSM1). However, none of these neuroendocrine markers are perfectly sensitive nor specific. Thus, we examined the utility of a calcium‐binding protein, secretagogin (SCGN), which is found mainly in neuroendocrine cells and neurons for diagnosing high‐grade neuroendocrine carcinomas (NECs) of the lung.Materials and MethodsA total of 71 pulmonary NEC resection specimens were collected from the pathology archive between 2005 and 2021, including 18 SCLCs, 13 combined‐SCLCs (c‐SCLCs), 23 LCNECs, and 17 combined‐LCNECs (c‐LCNECs). Immunohistochemical stains with SCGN were done and compared with synaptophysin, chromogranin A, CD56, and INSM1. The stains were evaluated by Allred scoring system which combined staining intensity and proportion of positive tumor cells. Both intensity score (none =0, mild =1, moderate =2, strong =3) and proportion score (0% =0, <1% =1, 1‐10% =2, 11‐33% =3, 34‐66% =4, 67‐100% =5) were summed up to give a final Allred score between 0 and 8. At least mild staining intensity in at least 1% of the cells was considered positive (Allred score greater than or equal to 2). Clinicopathological parameters such as age, sex, smoking history, tumor size, and tumor stage were also retrieved from the medical chart. Paired T‐test (two‐sided) was used to compare the Allred scores and positive proportions of each stain. Fisher’s exact test was used for categorical variables. A P‐value less than 0.05 was considered statistically significant.ResultsBioinformatic study showed the specific SCGN expression in neuroendocrine cells and NECs. No significant difference was observed among the four NEC categories regarding age, sex, tumor size, and staging. All patients had a history of smoking. SCGN showed diffuse nuclear and cytoplasmic staining in NECs with intra‐tumoral heterogenicity (Figure 1). The non‐neuroendocrine components were negative for SCGN. Sensitivity of SCGN was no better than the other established neuroendocrine markers based on Allred score for all cases combined or LCNECs/c‐LCNECs only. However, the sensitivity of SCGN (71%) was slightly higher than chromogranin A (68%) based on Allred score for SCLCs/c‐SCLCs only. The average percentage of SCGN positive tumor cells was 8% higher than chromogranin A (22% versus 14%, P=0.0332) in all NECs and 18% higher (32% for SCGN versus 13% for chromogranin A, P=0.0054) for SCLC and c‐SCLC cases only.ConclusionSCGN had the lowest sensitivity (55%) among the markers. However, it performed better than chromogranin A (71% versus 68%) in the diagnosis of SCLCs and c‐SCLCs. Its positive tumor cell percentage was significantly higher than chromogranin A in SCLCs and c‐SCLCs (18% higher, P=0.0054). The above data showed that SCGN could be used as a supplemental neuroendocrine marker to diagnose small cell carcinoma.

Melanoma Inhibitory Activity and Melanoma Inhibitory Activity 2 as Novel Immunohistochemical Markers of Oral Epithelial Dysplasia.

Oral potentially malignant disorders are associated with the development of oral squamous cell carcinoma (OSCC). Most OSCCs are diagnosed via histopathology as oral epithelial dysplasia (OED), but the histologic diagnostic criteria remain non-uniform. Accordingly, the establishment of a diagnostic marker to assist in diagnosis could contribute towards cancer prevention. Melanoma inhibitory activity (MIA) and MIA2 are involved in tumor growth, invasion, and lymph node metastasis in various malignancies. The purpose of this study was to clarify the usefulness of MIA and MIA2 as diagnostic markers of oral mucosal lesions. The expression of MIA and MIA2 was analyzed immunohistochemically in 100 specimens (10 specimens with normal oral mucosa (NOM) and 30 specimens each with low-grade epithelial dysplasia (LED), high-grade epithelial dysplasia (HED), and OSCC). Immunohistochemical results were evaluated based on the Allred scoring system. Cytoplasmic expression of MIA and MIA2 increased in the order of LED, HED, and OSCC. All NOM specimens were negative for cytoplasmic expression. Significant differences were observed between the groups (NOM vs. HED, p < 0.05, NOM vs. OSCC, p < 0.001). These results demonstrate that MIA and MIA2 are expressed in the oral mucosa within early neoplastic lesions and suggest that MIA and MIA2 are useful novel immunohistochemical markers for discriminating between normal tissue and OED.

Amongst Women Stratified to Receive Endocrine Therapy on the Basis of Their Tumor Estrogen and Progesterone Receptor Levels, Those with Higher Tumor Progesterone Receptor Levels Had a Better Outcome Than Those with Lower Levels of Tumor Progesterone Receptor.

Simple SummaryBreast cancer is the most commonly diagnosed cancer and the leading cause of cancer death of women worldwide. Several cut-points for estrogen receptor (ER) and progesterone receptor (PgR) have been proposed as predictive effects of hormone therapy; while the cut-off values were inconsistent. The aim of our retrospective study was to propose better prognostic cut-off levels for ER and PgR, and their effects on breast cancer-specific survival (BCSS) and disease-free survival (DFS) over 5 and 10 years were evaluated in 1807 eligible patients. Subgroups were generated based on ER and PgR expression percentage and scoring from the Allred scoring system (Allred scores). After comparing the hazard ratios (event rates in each group to reference group) of BCSS and DFS using multivariate analyses, our results suggested that patients with PgR expression ≤50% or Allred score ≤5 revealed a poor prognosis and should be paid more attention during follow-up.Background: To realize the association between stratified expression levels of ER and PgR and long-term prognosis of breast cancer patients who received adjuvant hormone therapy, this study aimed to propose better prognostic cut-off levels for estrogen receptor (ER) and progesterone receptor (PgR). Methods: Patients who received adjuvant hormone therapy after surgical intervention were selected. The ER and PgR status and their effects on breast cancer-specific survival (BCSS) and disease-free survival (DFS) over 5 and 10 years were evaluated. Next, subgroups were generated based on ER and PgR expression percentage and Allred scores. Survival curves were constructed using the Kaplan–Meier method. Results: ER and PgR expression were significantly associated with better prognosis in 5 years, whereas only PgR expression was significantly associated during the 10-year follow-up. The optimal cut-off values for better 5-year BCSS were ER > 50%; ER Allred score > 7; PgR ≥ 1%; or PgR Allred score ≥ 3; the corresponding values for DFS were ER > 40%; ER Allred score > 6; PgR > 10%; or PgR Allred score ≥ 3. In the long-term follow-up, PgR of > 50% or Allred score of > 5 carriers revealed a better prognosis of both BCSS and DFS. Conclusion: Patients with a PgR expression > 50% or an Allred score > 5 exhibited better 10-year BCSS and DFS.

Researches on the Correlation Between Estrogen and Progesterone Receptors Expression and Disease-Free Survival of Endometrial Cancer.

ObjectiveIn this study, 345 patients with endometrial carcinoma (EC) were selected to investigate the correlation between ER/PR status and the EC disease-free survival (DFS) rate.MethodsThe intensity and proportion of tumor cell expression of estrogen receptors and progesterone receptors (ER/PR) status of 345 postoperative tumor specimens in ECs were independently assessed semi-quantitatively by two pathologists using immunohistochemistry, the summed score ranged from 0 to 8 points was worked out by adding proportion score and intensity score based on the breast cancer hormone receptor immunohistochemical Allred scoring system. The association between DFS in ECs and ER/PR expression (intensity, proportion and summed score) was assessed using Cox regression analysis. Gene expression data were obtained from The Cancer Genome Atlas research network (TCGA).ResultsAccording to inclusion criteria, 201 type I and 144 type II EC patients were enrolled in this study. In the univariate analysis of type I endometrial carcinoma, the intensity, proportion and summed score of ER/PR status were significantly correlated with DFS. After adjusting for factors known to significantly impact survival, the influence of ER/PR status on DFS is generally decreased but the correlation is still significant. In the univariate analysis of type II endometrial carcinoma, the intensity, proportion and summed score of ER/PR status were significantly correlated with DFS. After adjusting for factors known to significantly impact survival, the influence of ER status on DFS is generally decreased, but the correlation is still significant, the effect of PR expression on DFS is not statistically significant.ConclusionHigher ER/PR expression status was associated with better DFS in patients with type I endometrial cancer after adjusting for known factors that significantly affect survival. In patients with type II endometrial cancer, patients with positive ER expression were significantly associated with better DFS. However, the effect of PR expression on DFS was not statistically significant.

Immunohistochemical expression of p16INK4A in the lesions of uterine cervix

Background: Cervical cancer is the major cause of cancer deaths among women. Globally, around 5,70,000 new cases of cervical cancer and 3,11,000 deaths occurred in the year 2018. In India, Cervical cancer is a leading cause of cancer related mortality among women and the number of deaths is 60,000 per year among 97,000 diagnosed patients, especially those from lower socioeconomic group. Human Papilloma Virus (HPV) plays a crucial role in causing cervical dysplasia. This is done by upregulating p16INK4A, a cyclin dependent kinase inhibitor through interaction with cellular regulatory proteins. Hence p16INK4A can be used as a biomarker, since it is directly related variable for the presence of HPV. This study was conducted to evaluate the expression of p16INK4A in benign, premalignant and malignant cervical lesions and to assess its utility in diagnosing and grading cervical lesions.&#x0D; Methods: A total of 80 cervical specimens categorized histopathologically into nonspecific cervicitis, low grade squamous intraepithelial neoplasia (LSIL), high grade squamous intraepithelial neoplasia (HSIL) and squamous cell carcinoma cervix were included in this prospective study of one-year duration. Immunohistochemical study of p16INK4A were interpreted qualitatively and semi-quantitatively by Allred scoring system (0 to 8 points) which measures the proportion of stained cells and intensity of staining of cells. The collected data were statistically analyzed by ANOVA and chi square test.&#x0D; Result: Qualitative method showed absence of p16INK4A expression in all nonspecific cervicitis. 16.7% (2/12) LSIL, 100% (12/12) HSIL and 100% (28/28) squamous cell carcinoma cases showed p16INK4A positivity. Allred scoring of p16INK4A showed 66% (8/12) HSIL and 85.7% (24/28) squamous cell carcinoma cases with score 3 positivity. Hence high-grade lesions showed higher expression of this marker.&#x0D; Conclusion: IHC expression of p16INK4A showed increasing degree of expression from benign to premalignant and malignant lesions suggesting its diagnostic and prognostic value in the cervical cancer management

151P - Gene expression profiling for a better understanding of gastric cancer: From the perspective of metabolic rearrangement

BackgroundMetabolic rearrangement has been shown to be an important characteristic for stomach adenocarcinomas. Here we aimed to improve our understanding of the tumorigenesis of gastric cancer by means of gene and protein expression profile analysis with a focus on metabolic genes and pathways. MethodsArray-based gene expression profiling of fresh frozen cancer tissues and adjacent normal tissues were obtained from 8 patients with gastric cancer at early stage by using Affymetrix oligonucleotide microarray. Assays targeted 179 unique genes related to cancer metabolism. The raw expression data were normalized using nSolver Analysis Software 3.0 and a dataset of gene expression ratios for GC vs. controls was generated. The p values were calculated using a paired t-test, and the threshold for up- and down-regulated genes was set at p value < 0.05. The protein expression of the dysregualted genes were detected by immunohistochemistry (IHC) in the formalin fixed paraffin embedded tissue blocks. The signal was quantified by the Allred score system which represented the estimated intensity and proportion of positive-staining cells. ResultsOur microarray results showed increased expression of 20 metabolic genes and decreased expression of 6 metabolic genes in all cases of gastric cancer at early stage. Besides of the undetected NOX4, the protein levels of all the others dysregualted genes, detected by IHC, showed consistently results. Half of all dysregulated genes (AKT2, EGLN3, G6PD, GLS, HIF1A, HK2, HRAS, MAP2K1, NTRK3, PGK1, PLCG1, RET and RPS6KB1) are implicated in Carbon Metabolism, a pivotal metabolic approach involving in nucleic acid biosynthesis. Five genes (ARNT, EGLN1, EGLN3, HIF1A and NOX4) are molecules implicated in hypoxia signaling. Besides, the upregulated HIF1A is a cancer metabolism driver, which means that HIF1A may induce the oncogenesis of gastric cancer. ConclusionsThe gastric mucosa in gastric cancer at early stage is characterized by dysregulated expression of a limited repertoire of metabolic genes. The nature of the corresponding metabolic rearrangement and pathways may help guide further investigations into its etiology. Legal entity responsible for the studyFudan University Shanghai Cancer Center. FundingNational Human Genetic Resources Sharing Service Platform (2005DKA21300), National Natural Science Foundation of China (81602078),. DisclosureAll authors have declared no conflicts of interest.

The Quantitative ER Immunohistochemical Analysis in Breast Cancer: Detecting the 3 + 0, 4 + 0, and 5 + 0 Allred Score Cases.

Background and objectives: The currently used immunohistochemical approach in determining the estrogen receptor (ER) positivity of breast cancers (BCs) is inherently subjective and additionally limited by its semi-quantitative nature. The application of software in the analysis of digitized slide images may overcome some of these limitations. However, the utilization of such an approach requires that the entire staining procedure is standardized. Background and objectives: We aimed to establish a procedure for the photometric and morphometric analysis of BC immunohistochemical parameters that can possibly be used for a diagnostic purpose that is in line with the current semi-quantitative scoring system. Materials and Methods: Semi-quantitative analysis of ER-stained tissue sections was performed following the Allred scoring system guidelines. The quantitative analysis was performed in ImageJ software after color deconvolution. The quantitative analysis of 66 cases of invasive lobular BC included: Percent of ER-positive cells, average nuclear coloration intensity, and the quantitative ER score. The percent of ER-positive tumor cells was counted using a standard grid overlay, while optical density (0.0–1.0) was measured within each nucleus at the grid points. Results: A statistical analysis revealed a significant positive correlation (r = 0.886, p < 0.001) between the subjective semi-quantitative and quantitative ER scores, with a large effect size (d = 3.8215). We observed strong statistically significant correlations between individual parameters of the total ER score, percentage of ER-positive nuclei, and color intensity, obtained by the two independent methods. Conclusions: Additionally, besides excluding subjectivity, the up to now unreported cases of 3 + 0, 4 + 0, and 5 + 0 Allred scores were detected only by the application of the proposed quantitative approach.

Evaluation of deslorelin implant on subsequent mammary tumors of rats (Rattus norvegicus)

ABSTRACT Background Mammary fibroadenomas are one of the most common tumors of female companion rats (Rattus norvegicus forma domestica). The objectives of this study were to determine if subcutaneous administration of a deslorelin implant following excision of fibroadenomas can prevent or delay development of additional mammary tumors and increase survival in companion rats. Methods Female intact client-owned rats with benign mammary tumors were divided into three groups: no implant (n = 10), placebo implant (n = 10), or 4.7 mg deslorelin implant (n = 10) placed within 2 months of tumor excision. Rats were monitored for subsequent mammary tumors for 10 months following treatment. Expression of estrogen α, progesterone, prolactin, and androgen receptors stained by immunohistochemistry in primary masses of rats included in the deslorelin-treated group was evaluated using the Allred scoring system. Results In the control non-implanted group, four of the 10 rats developed another mammary tumor, including one anaplastic carcinoma. In the control placebo group, five of the 10 rats developed another mammary tumor, including one ductal carcinoma. In the deslorelin-treated group, three of 10 rats developed another mammary tumor, including one adenocarcinoma. Median time between surgery and new mass detection did not differ significantly among groups (55 days in the deslorelin group vs. 77 days in the control placebo group, P = 0.25). Median survival times after surgery did not differ significantly among groups. No correlation was noted between receptor expression and response to treatment with deslorelin. Conclusions and clinical relevance Deslorelin implants placed within 2 months of benign mammary tumor surgical excision were not associated with a decreased risk of developing subsequent mammary tumors, nor with an increased survival in female rats. Further studies are needed to define useful adjunct therapy to surgery.

Abstract 1074: Expression of p39 and Rb in lung cancer and its correlation with clinicopathologic parameters

Abstract Lung cancer is characterized by its aggressiveness and proclivity for early metastasis. A predictor of metastasis of NSCLC rely on post-resection evaluation of tumor histology, which is a severe limitation since only 15% of the patients are diagnosed with resectable disease. Hence, there is a need to characterize biomarkers with metastasis-predicting value in pre-resection small biopsy specimens. Rb is a tumor suppressor inactivated due to hyper-phosphorylation in NSCLC. Our preliminary data relates the Rb S249 phosphorylation to an epithelial-to-mesenchymal transition (EMT), a trait strongly associated to metastasis and increase in CDK5 activator p39. Here, we examined the prognostic significance of Rb S249 and p39 as biomarkers of metastasis staging in non-small cell lung cancer. Our hypothesis is that Rb hyper-phosphorylation in S249 can have prognostic value by being associated with a metastatic phenotype and EMT. To determine p39 and Rb phosphorylation in lung cancer, immunohistochemistry analysis was performed on two lung cancer microarrays. Three pathologists observed the microarrays and scored them using the all-red scoring system. P39 and Rb phosphorylation in S249 were more present in lung tumors than adjacent normal tissue. P39 positive expression had a positive correlation with staging, lymph node involvement, and metastasis. Rb phosphorylation in residue S249 had a positive correlation with histological grading. Using general linear model, we determined that staging can be predicted when combining phosphorylation of Rb S249 and p39. Our analysis suggests that p39 and Rb S249 are likely to be diagnostic biomarkers for lung cancer metastasis. Citation Format: Jaileene Perez-Morales, Stephanie Rivera, Bryan Torres, Xavier Rodriguez, Angel Isidro, Pedro Santiago-Cardona. Expression of p39 and Rb in lung cancer and its correlation with clinicopathologic parameters [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1074.

Tissue expression of retinoic acid receptor alpha and CRABP2 in metastatic nephroblastomas

BackgroundNephroblastoma or Wilms tumor is the most frequent kidney cancer in children and accounts for 98% of kidney tumors in this age group. Despite favorable prognosis, a subgroup of these patients progresses to recurrence and death. The retinoic acid (RA) pathway plays a role in the chemoprevention and treatment of tumors due to its effects on cell differentiation and its antiproliferative, anti-oxidant, and pro-apoptotic activities. Reports describe abnormal cellular retinoic acid-binding protein 2 (CRABP2) expression in neoplasms and its correlation with prognostic factors and clinical and pathological characteristics. The aim of this study was to evaluate the immunohistochemical expression of retinoic acid receptor alpha (RARA) and CRABP2 in paraffin-embedded samples of nephroblastomas via semiquantitative and quantitative analyses and to correlate this expression with prognostic factors.MethodsSeventy-seven cases of nephroblastomas were selected from pediatric oncology services. The respective medical records and surgical specimens were reviewed. Three representative tumor samples and one non-tumor renal tissue sample were selected for the preparation of tissue microarrays (TMA). The Allred scoring system was used for semiquantitative immunohistochemical analyses, whereas a morphometric analysis of the stained area was employed for quantitative evaluation. The nonparametric Mann-Whitney test was used for comparisons between two groups, while the nonparametric Kruskal-Wallis test was used to compare three or more groups.ResultsImmunopositivity for RARA and CRABP2 was observed in both the nucleus and cytoplasm. All histological components of the nephroblastoma (blastema, epithelium, and stroma) were positive for both markers. RARA, based on semiquantitative analyses, and CRABP2, bases on quantitative analyses, exhibited increased immunohistochemical expression in patients with metastasis, with p values of 0.0247 and 0.0128, respectively. These findings were similar to the results of the quantitative analysis of RARA expression, showing greater immunopositivity in tumor samples of patients subjected to pre-surgical chemotherapy. No significant correlation was found with the other variables studied, such as disease stage, anaplasia, risk group, histological type, nodal involvement, and clinical evolution.ConclusionsSemiquantitative and quantitative analyses of the markers RARA and CRABP2 indicate their potential as biomarkers for tumor progression and their participation in nephroblastoma tumorigenesis.

Evaluation of Immunohistochemical Profile of Breast Cancer for Prognostics and Therapeutic Use.

Introduction:Breast cancer is leading cancer in women, and the incidence of breast cancer in India is on the rise. The most common histologic type of breast cancer is infiltrating ductal carcinoma. Prognostic and predictive factors are used in the management of breast cancer. Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2/neu) are immunohistochemical markers of prognosis as well as predictors of response to therapy.Aims and Objectives:The study was conducted to evaluate ER, PR, and HER2/neu expressions in invasive ductal carcinomas of the breast by immunohistochemistry, to explore the correlation of these markers to each other and to various clinicopathological parameters: age of the patient, histological grade, tumor size, and lymph node metastasis.Materials and Methods:This prospective study was conducted on 100 cases of infiltrating ductal carcinoma. Slides were prepared from blocks containing cancer tissue, and immunohistochemical staining was done for ER, PR, and HER2/neu expressions. Interpretation of expressions was done using Allred scoring system for ER/PR and the American Society of Clinical Oncology/College of American Pathologists guidelines for HER2/neu. Statistical analysis was performed to determine the statistical significance by applying Chi-square test.Results:Majority of tumors were ER and PR positive and HER2/neu negative. ER and PR correlated significantly with age, tumor size, and tumor grade; whereas, HER2/neu correlated significantly with tumor size only. No association was seen with axillary lymph node metastasis. ER and PR expression correlated with each other, but none correlated with HER2/neu.Conclusions:As the majority of the tumors are ER, PR positive and since ER and PR correlate with each other as well as with age, tumor size, and grade. Therefore, routine assessment of hormone receptors is recommended for prognostic and therapeutic information in breast cancer cases.

Breast cancer subtype discrimination using standardized 4-IHC and digital image analysis.

Breast cancer is a heterogeneous disease. Surrogate classification of intrinsic subtypes of invasive carcinomas by combined immunohistochemistry for estrogen receptor (ER), progesterone receptor (PR), HER2, and Ki67 (4-IHC) has increased steadily since the 2011 St Gallen symposium, due to its rapid subtyping of tumors at a reasonable cost. An important step in improving 4-IHC reproducibility and reliability will be to provide reference values from the routine use of standardized 4-IHC followed by image analysis. The aims of the current study were (1) to analyze invasive breast carcinomas using standardized 4-IHC and quantitative image analysis and (2) to compare the results obtained in the classification of biological subtypes using current Ki67 and PR threshold values proposed by different authors to sub-classifying the luminal A-like and the luminal B-like (HER2-negative) subtypes. Five hundred twenty-one tumors were analyzed by standardized immunohistochemistry, with automatic image analysis, and HER2 FISH technique. Positivity for ER was found in 82.7% and for PR in 70.1% of cases. Using the Allred scoring system, hormone receptor results showed a bimodal distribution, particularly for ER. HER2 positivity was found in 15.7% of cases, and the mean Ki67 score was 32.3%. Using the most recently proposed surrogate definitions for the classification of luminal breast cancer subtypes, the percentages of different subtypes that we found were similar to those published with genomic platforms: 40.7% luminal A-like, 32.4% luminal B-like/HER2-negative, 9.8% luminal B-like/HER2-positive, 6.0% HER2-positive, and 11.1% triple negative. Standardized 4-IHC with automatic image analysis constitutes a low-cost method for surrogate definitions of biological subtypes of breast cancer that delivers accurate results in a day.

Differential expression of nucleolin in colon adenoma and adenocarcinoma

Abstract Objective The aim of the study was to investigate the discrepancy in nucleolin expression between colon adenoma and colon adenocarcinoma, explore the role of nucleolin expression in the carcinogenesis of colon adenocarcinoma, and determine the correlation of the nucleolin expression level with histological grade in colon adenocarcinoma. Methods In total, 80 cases of colon adenocarcinoma with cancer-adjacent colon mucosa and 60 cases of colon adenomas were examined by immunohistochemistry using an antibody against nucleolin. Nucleolin expression levels in these groups were compared. The correlation between the nucleolin expression level and grade of colon adenocarcinoma was analyzed. Results Nucleolin expression is located in the nuclei of colon adenocarcinoma, colon adenoma, and cancer-adjacent colon mucosa tissues with different intensities. A semiquantitative evaluation using the Allred scoring system showed that the nucleolin immunostaining score in colon adenocarcinoma (7.8 ± 0.1) was significantly higher than those in colon adenoma (6.3 ± 0.2) and cancer-adjacent colon mucosa (5.4 ± 0.1; P &lt; 0.01). The nucleolin immunostaining score in colon adenoma was significantly higher than that in cancer-adjacent colon mucosa (P &lt; 0.01). Nucleolin expression levels in well-differentiated and moderately differentiated adenocarcinoma (6.8 ± 0.2) were significantly lower than those in poorly differentiated adenocarcinoma (8.0 ± 0.1; P &lt; 0.01). Conclusion Increased nucleolin expression may play an important role in the process of malignant transformation of colon adenocarcinoma and predicts a poor prognosis.