Positive Staining Of Tumor Cells Research Articles

High expression of NF-κB inducing kinase in the bulge region of hair follicle induces tumor

The bulge region, a reservoir of multipotent stem cells, is possibly responsible for tumorigenesis. NF-κB-inducing kinase (NIK) is a kinase involved in the activation of the noncanonical NF-κB pathway and exhibits positive staining in tumor cells. However, whether high expression of NIK can result in tumorigenesis has not been reported in published papers. By establishing Nik-coe (Nik-stopF/F crossed with Chat-cre) and Nik-soe (Nik-stopF/F crossed with Sox9-cre) mice, we found that overexpression of Nik in the bulge region of hair follicles induced hair follicle loss and tumorigenesis. Furthermore, RNA sequencing, proteomic and phosphopeptide analyses revealed that multiple cancer pathways are involved in tumor formation. Taken together, these findings indicate that constitutive activation of Nik in the bulge region induces tumorigenesis.

Estrogen Receptor Status by Immunohistochemistry Is Superior to the Ligand-Binding Assay for Predicting Response to Adjuvant Endocrine Therapy in Breast Cancer.

Immunohistochemistry (IHC) is a newer technique for assessing the estrogen receptor (ER) status of breast cancers, with the potential to overcome many of the shortcomings associated with the traditional ligand-binding assay (LBA). The purpose of this study was to evaluate the ability of ER status determination by IHC, compared with LBA, to predict clinical outcome-especially response to adjuvant endocrine therapy-in a large number of patients with long-term clinical follow-up. ER status was evaluated in 1,982 primary breast cancers by IHC on formalin-fixed paraffin-embedded tissue sections, using antibody 6F11 and standard methodology. Slides were scored on a scale representing the estimated proportion and intensity of positive-staining tumor cells (range, 0 to 8). Results were compared with ER values obtained by the LBA in the same tumors and to clinical outcome. An IHC score of greater than 2 (corresponding to as few as 1% to 10% weakly positive cells) was used to define ER positivity on the basis of a univariate cut-point analysis of all possible scores and disease-free survival (DFS) in patients receiving any adjuvant endocrine therapy. Using this definition, 71% of all tumors were determined to be ER-positive by IHC, and the level of agreement with the LBA was 86%. In multivariate analyses of patients receiving adjuvant endocrine therapy alone, ER status determined by IHC was better than that determined by the LBA at predicting improved DFS (hazard ratios/P = 0.474/.0008 and 0.707/.3214, respectively) and equivalent at predicting overall survival (0.379/.0001 and 0.381/.0003, respectively). IHC is superior to the LBA for assessing ER status in primary breast cancer because it is easier, safer, and less expensive, and has an equivalent or better ability to predict response to adjuvant endocrine therapy.

Abstract 1790: Herceptin-resistant breast cancer cells exhibits distinct sensitivity to lapatinib in in vivo tumor xenograft models

Abstract Introduction: HER2-targeted therapies, including the monoclonal antibody (Ab) herceptin and the tyrosine kinase inhibitor (TKI) lapatinib, have been commonly used to treat HER2-positive breast cancer (BC). However, resistance to herceptin and/or lapatinib frequently occurs and currently represents a significant clinical problem. We previously demonstrated that both HER3- and IGF-1 receptor (IGF-1R)-initiated signaling played crucial roles in the development of herceptin resistance. Further studies showed that our herceptin-resistant SKBR3-pool2 and BT474-HR20 sublines, as compared their parental SKBR3 and BT474 cells, respectively, also exhibited refractoriness to lapatinib in vitro. However, the in vivo efficacy of lapatinib against the herceptin-resistant BC models (SKBR3-pool2 and BT474-HR20) remains unclear. Methods: Tumor xenograft models were established via subcutaneous inoculation of SKBR3-pool2 or BT474-HR20 cells into athymic nude mice. Tumor formation was assessed by palpation and measured with fine calipers twice a week. The tumor-bearing mice were treated with vehicle (DMSO) or lapatinib (80 mg/kg) via intraperitoneal injection daily for three weeks. Immunohistochemistry (IHC) assays were performed to assess the expression and activation of proteins. Two individuals independently read and quantified all IHC slides by considering both the intensity of immunostaining and the percentage of positive-staining tumor cells. Results: Treatment with lapatinib profoundly inhibited growth of the tumors established with SKBR3-pool2 cells. In contrast, lapatinib slightly, but not significantly, reduced tumor growth in BT474-HR20 cells-derived xenograft models. IHC analyses of the tumors obtained from both SKBR3-pool2 and BT474-HR20 xenografts showed that lapatinib treatment had little effect on the expression of HER3 and the protein phosphatase PPP3CB. Lapatinib also did not alter the levels of phosphorylated Akt (p-Akt) and FOXO3a (p-FOXO3a) in vivo. Interestingly, in BT474-HR20-derived tumors, lapatinib treatment dramatically enhanced expression of insulin receptor substrate-1 (IRS1) and increased the levels of phosphorylated HER3 (p-HER3). However, such phenomena were not observed in the tumors established with SKBR3-pool2 cells. Conclusion: While the tumors derived from BT474-HR20 cells retain their resistant phenotype to lapatinib, SKBR3-pool2-tumors are highly sensitive to lapatinib treatment. It seemed that lapatinib-induced upregulation of IRS1 and activation of HER3, evidenced by increased p-HER3, contributed to the inefficacy of lapatinib against BT474-HR20-tumors in vivo. Keywords: HER3, IRS1, treatment resistance, HER2-targeted therapy, breast cancer Citation Format: Sanbao Ruan, Hao Liu, Hui Lyu, Congcong Tan, Bolin Liu. Herceptin-resistant breast cancer cells exhibits distinct sensitivity to lapatinib in in vivo tumor xenograft models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1790.

Mucinous adenocarcinoma of Prostate (MAOP), a rare variant of prostate adenocarcinoma: a case report and literature review

A prostatectomy specimen was received from a 72-year-old male with a previous needle core biopsy diagnosis of prostatic adenocarcinoma, Gleason score 3+4=7. Microscopic examination of the prostatectomy specimen demonstrated single and fused neoplastic glands floating in copious amounts of extracellular mucin representing >50% of the tumour volume. There was a minor component of non-mucinous conventional adenocarcinoma in the background. Immunhistochemistry NKX 3.1 and PSA showed positive staining of tumour cells. Clinical, radiological or serological evidence of carcinoma of another site was not revealed. A diagnosis of MAOP, Gleason score 4+3 =7 (WHO/ISUP Grade Group 3) with a tertiary component of Gleason pattern 5 comprising <1% of total tumour volume was made. MAOP is a rare variant, comprising 0.38–0.43% of prostatic carcinoma. It is characterised by tumour cells floating in >25% of extracellular mucin excluding any intraluminal mucin. The diagnosis should only be made in resection specimens, and following exclusion of secondary deposits of extra-prostatic origin of mucinous adenocarcinoma by clinical-radiological correlation and with confirmatory immunohistochemistry. Grading and scoring for MAOP should be performed according to the growth pattern similar to conventional adenocarcinoma. The prognosis appears to be similar to non-mucinous prostatic adenocarcinoma.

GIANT PRIMARY COLLOID CARCINOMA OF THE LUNG

SESSION TITLE: Monday Medical Student/Resident Case Report Posters SESSION TYPE: Med Student/Res Case Rep Postr PRESENTED ON: 10/21/2019 02:30 PM - 03:15 PM INTRODUCTION: Colloid carcinomas of the lung are a very rare subset of lung adenocarcinomas. Historically, these neoplasms have been called mucinous cystic tumors and cystic mucinous adenocarcinomas. These tumors can grow very large and appear gelatinous and mucoid. We report a case of an extremely rare giant colloid carcinoma in a relatively asymptomatic patient. CASE PRESENTATION: The patient was a 55-year-old male with chronic kidney disease who was referred to our thoracic surgery service after a huge right lung mass was incidentally found during evaluation for renal transplantation. On MRI, it measured 11.8 x 9.4 x 13.1 cm and occupied most of the right chest cavity. The physical exam surprisingly demonstrated no abnormalities other than diminished breath sounds in the right lower lung base. Prior to surgery, bronchoscopy demonstrated endobronchial tumor extending from the right lower lobe into the intermedius bronchus. It was felt that the patient would require lower and middle lobectomy to obtain adequate tumor clearance. A hilar and lymph node dissection was started using the da Vinci robotic-assisted surgery system to see if adequate vascular control can be obtained in the hilum. After limited success a right thoracotomy was performed in order to complete the dissection and removal of the massive tumor. He made a satisfactory post-operative recovery and was discharged 10 days post-operatively doing well. Five month follow-up visit indicates that he is continuing to improve and currently waiting oncology clearance for consideration for kidney transplantation. The tumor was evaluated to have clear margins and measured 15.5 x 12.2 x 6.5 cm. Final pathologic diagnosis was a 15.5 cm colloid carcinoma of the lung with no evidence of lymph node metastasis. IHC staining revealed CK7 and CDX2 positive-staining tumor cells. DISCUSSION: Colloid carcinoma of the lung is a very rare diagnosis. These tumors are normally evident in breast tissue or the gastrointestinal tract. Because of the higher prevalence in tissues other than the lung, it is important to make sure the lung tumor is not a distant metastasis. In our case, it was important to rule out distant metastasis because of the patient’s potential renal transplantation status. The tumor’s IHC staining was consistent with colloid carcinomas of the lung. To our knowledge, our case represents one of the largest colloid carcinomas of the lung ever documented. The largest tumor we were able to find in the literature measured 15 cm at its largest. Our patient’s tumor measures 15.5 cm at the largest diameter. Complete resection tumor resection with mediastinal lymph node staging is the recommended treatment. CONCLUSIONS: The patient presented with an incidental and rare gigantic lung tumor. Despite the size of the tumor, it was completely resected with a possible cure. Reference #1: Moran CA, Hochholzer L, Fishback N, Travis WD, Koss MN. Mucinous (so-called colloid) carcinomas of lung. Modern pathology: an official journal of the United States and Canadian Academy of Pathology, Inc. 1992 Nov;5(6):634-8. Reference #2: Gao ZH, Urbanski SJ. The spectrum of pulmonary mucinous cystic neoplasia: a clinicopathologic and immunohistochemical study of ten cases and review of the literature. American journal of clinical pathology. 2005 Jul 1;124(1):62-70. Reference #3: Masai K, Sakurai H, Suzuki S, Asakura K, Nakagawa K, Watanabe SI. Clinicopathological features of colloid adenocarcinoma of the lung: A report of six cases. Journal of surgical oncology. 2016 Aug;114(2):211-5. DISCLOSURES: No relevant relationships by John Hallsten, source=Web Response No relevant relationships by Wickii Vigneswaran, source=Web Response

Expression of Immunomodulatory Molecules PD-1, PD-L1, and PD-L2, and their Relationship With Clinicopathologic Characteristics in Endometrial Cancer.

Chemotherapeutic agents are not very effective in treating advanced endometrial cancers (ECs). Recent studies have demonstrated the immune evasion mechanism of tumors and possible remedies. Programmed cell death protein 1 (PD-1), programmed death ligand 1 (PD-L1), and programmed death ligand 2 (PD-L2) are immunomodulator molecules that have been the focus of research in lung cancer, melanoma, and renal cell cancer. However, there are few studies concerning EC. This retrospective study aimed to determine PD-1, PD-L1, and PD-L2 expression immunohistochemically in EC, and to study their correlation with clinicopathologic tumor characteristics. This study comprised 127 patients with EC. Anti PD-1, PD-L1, and PD-L2 antibodies were examined immunohistochemically on sections obtained from tissue microarray paraffin blocks. No staining with PD-1 in tumor cells was seen; however, we found positive staining in tumor cells at 36.2% with PD-L1 and 64.4% with PD-L2, and at 61.6% with PD-1, 36.2% with PD-L1, and 93.2% with PD-L2 in immune cells. When comparing staining and clinicopathologic findings, most of the PD-L1 negative tumors (both in tumor and immune cells) were FIGO Stage I, which was significantly higher than stage II-III-IV tumors (P<0.05). There was a statistically significant association between the FIGO grade and the PD-L1 score in immune cells (P=0.009), and between staining of PD-1, PD-L1, and PD-L2 and age (P=0.004, 0.013, and 0.043, respectively). Interaction between PD-1, PD-L1, and PD-L2 may be a potential target for immunotherapy in elderly and advanced stage EC patients.

Prognostic Role of S100A8 and S100A9 Protein Expressions in Non-small Cell Carcinoma of the Lung.

BackgroundS100A8 and S100A9 have been gaining recognition for modulating tumor growthand metastasis. This study aimed at evaluating the clinical significance of S100A8 and S100A9 innon-small cell lung cancer (NSCLC).MethodsWe analyzed the relationship between S100A8and S100A9 expressions, clinicopathological characteristics, and prognostic significance in tumorcells and peritumoral inflammatory cells.ResultsThe positive staining of S100A8 in tumorcells was significantly increased in male (p < .001), smoker (p = .034), surgical method other thanlobectomy (p = .024), squamous cell carcinoma (SQCC) (p < .001) and higher TNM stage (p = .022)compared with female, non-smoker, lobectomy, adenocarcinoma (ADC), and lower stage. Theproportion of tumor cells stained for S100A8 was related to histologic type (p < .001) and patientsex (p = .027). The proportion of inflammatory cells stained for S100A8 was correlated with patientage (p = .022), whereas the proportion of inflammatory cells stained for S100A9 was correlatedwith patient sex (p < .001) and smoking history (p = .031). Moreover, positive staining in tumorcells, more than 50% of the tumor cells stained and less than 30% of the inflammatory cellsstained for S100A8 and S100A9 suggested a tendency towards increased survivability in SQCCbut towards decreased survivability in ADC.ConclusionsS100A8 and S100A9 expressions might be potential prognostic markers in patients with NSCLC.

Plasmin(ogen) serves as a favorable biomarker for prediction of survival in advanced high-grade serous ovarian cancer.

In serous ovarian cancer, the clinical relevance of tumor cell-expressed plasmin(ogen) (PLG) has not yet been evaluated. Due to its proteolytic activity, plasmin supports tumorigenesis, however, angiostatin(-like) fragments, derived from PLG, can also function as potent anti-tumorigenic factors. In the present study, we assessed PLG protein expression in 103 cases of advanced high-grade serous ovarian cancer (FIGO III/IV) by immunohistochemistry (IHC). In 70/103 cases, positive staining of tumor cells was observed. In univariate Cox regression analysis, PLG staining was positively associated with prolonged overall survival (OS) [hazard ratio (HR)=0.59, p=0.026] of the patients. In multivariable analysis, PLG, together with residual tumor mass, remained a statistically significant independent prognostic marker (HR=0.49, p=0.009). In another small patient cohort (n=29), we assessed mRNA expression levels of PLG by quantitative PCR. Here, elevated PLG mRNA levels were also significantly associated with prolonged OS of patients (Kaplan-Meier analysis; p=0.001). This finding was validated by in silico analysis of a microarray data set (n=398) from The Cancer Genome Atlas (Kaplan-Meier analysis; p=0.031). In summary, these data indicate that elevated PLG expression represents a favorable prognostic biomarker in advanced (FIGO III/IV) high-grade serous ovarian cancer.

Abstract 4027: Recurrent gains of CD274 (PD-L1) and PDCD1LG2 (PD-L2) provide a genetic basis for PD-1 ligand expression in a subset of solid tumors

Abstract Background: A subset of malignant tumors are immunogenic but avoid immune surveillance through ectopic expression of the programmed cell death 1 ligands, PD-L1 and PD-L2, that bind PD-1 on effector T-cells to inhibit T-cell activation, a phenotype that is reversible with PD-1 blockade. Co-amplification and co-gain of CD274 (PD-L1) and PDCD1LG2 (PD-L2) on chromosome 9p24.1 provide a genetic basis for PD-1 ligand expression by Reed-Sternberg cells in classical Hodgkin lymphoma, a tumor highly sensitive to PD-1 blockade. We sought to determine whether copy number alterations of CD274 and PDCD1LG2 are a mechanism of PD-1 ligand expression in other tumor types. Methods: We surveyed data derived from a clinically-deployed next generation sequencing assay (OncoPanel) to identify tumors with evidence of selective copy gain of genes located at 9p24.1. Formalin-fixed paraffin-embedded biopsy specimens from available cases were reviewed for tumor content, immunostained for PD-L1, and analyzed with custom probes targeting the CD274 and PDCD1LG2 loci and a commercial probe targeting the centromere of chromosome 9 by flourescence in situ hybridization (FISH). Results: Review of NGS data revealed 171 of 7,070 clinical samples (2.4%) with evidence for selective copy gain of CD274 and/or PDCD1LG2. Tissue was available for analysis in 73 cases, of which 55 were successfully analyzed. Co-amplification and co-gain of CD274 and PDCD1LG2 were observed for 20 (36%) and 18 (33%) of cases, respectively. Polysomy 9 and disomy 9 were observed among 9 (16%) and 2 (4%) cases, respectively. Genetic abnormalities associated with chromosome 9 loss were observed among the remaining cases 6 (11%). PD-L1 IHC revealed positive staining of tumor cells in 36 (65%) cases. Among cases with co-amplification/gain of CD274 and PDCD1LG2, a subset (Co-Amp: 36%; Co-Gain: 28%) exhibited moderate to high PD-L1 expression in significant percentages of tumor cells (Tumor H-score &amp;gt; 50). Tumor types with high PD-L1 expression in the context of co-amplification/gain included carcinomas of the head and neck, bladder, and cervix. Conclusions: Review of over 7000 tumors analyzed by a clinically-deployed NGS platform identifies CD274 and PDCD1LG2 copy gain in a low percentage of a variety of solid tumor types, a subset of which has a corresponding increase of PD-L1 expression. These data suggest that copy gain of CD274 and PDCD1LG2 contributes to PD-1 ligand expression in a subset of solid tumors beyond classical Hodgkin lymphoma. Citation Format: Evisa Gjini, Christine Pak, Alyssa Kelly, Yuling Shi, Neal Lindeman, Frank Kuo, Gordon Freeman, Azra Ligon, Stephen Hodi, Margaret Shipp, Scott Rodig. Recurrent gains of CD274 (PD-L1) and PDCD1LG2 (PD-L2) provide a genetic basis for PD-1 ligand expression in a subset of solid tumors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4027.

Immune checkpoints programmed death 1 ligand 1 and cytotoxic T lymphocyte associated molecule 4 in gastric adenocarcinoma

ABSTRACTRemarkable efficacy of immune checkpoint inhibition has been reported for several types of solid tumors and early studies in gastric adenocarcinoma are promising. A detailed knowledge about the natural biology of immune checkpoints in gastric adenocarcinoma is essential for clinical and translational evaluation of these drugs. This study is a comprehensive analysis of cytotoxic T lymphocyte associated molecule 4 (CTLA-4) and programmed death 1 ligand 1 (PD-L1) expression in gastric adenocarcinoma. PD-L1 and CTLA-4 were stained on tumor sections of 127 Caucasian patients with gastric adenocarcinoma by immunohistochemistry (IHC) and somatic mutation profiling was performed using targeted next-generation sequencing. Expression of PD-L1 and CTLA-4 on lymphocytes in tumor sections, tumor-draining lymph nodes (TDLN) and peripheral blood were studied by flow-cytometry and immune-fluorescence microscopy in an additional cohort. PD-L1 and CTLA-4 were expressed in 44.9% (57/127) and 86.6% (110/127) of the analyzed gastric adenocarcinoma samples, respectively. Positive tumor cell staining for PD-L1 or CTLA-4 was associated with inferior overall survival. Somatic mutational analysis did not reveal a correlation to expression of PD-L1 or CTLA-4 on tumor cells. Expression of PD-1 (52.2%), PD-L1 (42.2%) and CTLA-4 (1.6%) on tumor infiltrating T cells was significantly elevated compared to peripheral blood. Of note, PD-1 and PD-L1 were expressed far higher by tumor-infiltrating lymphocytes than CTLA-4. In conclusion, specific immune checkpoint-inhibitors should be evaluated in this disease and the combination with molecular targeted therapies might be of benefit. An extensive immune monitoring should accompany these studies to better understand their mode of action in the tumor microenvironment.

A51: Immunologic microenvironment and some molecular biological characteristics of breast cancer

The purpose of the study was to assess parameters of local cellular immunity, expression of apoptosis-controlling proteins and some other receptors in various molecular subtypes of breast cancer (BC) with different clinical course and prognosis. Materials and methods 100 BC patients aged 30–76 years (59 ± 3.4) were recruited. Tissues of tumor and peritumoral area were homogenized by MediMachine, subset contents of T, B, NK-lymphocytes were estimated by flow cytometer FACS CantoII. Percentage of lymphocytes expressing CD3, CD4, CD8, CD19, CD16/56, were counted from total amount of CD45+ lymphocytes. Sections of paraffin-embedded blocks were studied by immunohistochemical method with polyvalent HRP-DAB detection system. Staining was performed using Thermo Scientific autostainer. Tumor was considered p53-positive when >25% of tumor cell nuclei were positively stained, and bcl-2-positive when >25% of cells showed specific cytoplasmic staining. Expression of E-cadherin was assessed by the number of cells with positive membrane expression of this marker, taking into account intensity of the reaction. Expression of topoisomerase 2α and androgen receptors was assessed by the number of tumor cells with positive nuclear expression of these markers; number of stained nuclei per 100 nuclei in 3 fields of view was counted. Results Some differences characterizing tumor cells of molecular breast cancer subtypes were found. Tumor tissue contained higher amount of T-lymphocytes than blood (85.9 ± 2.36% vs 60.1 ± 4.5%, P p53 overexpression was equally often recorded in luminal B and Her2+ subtypes of BC and slightly less frequently in triple-negative subtype. Accumulation of p53 was 2 times less frequently detected in luminal A subtype in comparison with triple -negative cancer and 2.5 and 2.7 times less frequently in comparison with luminal B and Her2+ BC, respectively. Study of bcl-2 expression showed reducing the frequency of positive tumor cell staining in dependence on the receptor status of tumor: positive cases were several times less frequent in triple-negative and Her2+ subtypes in comparison with luminal A and B subtypes. Luminal A subtype was characterized by the minimal p53, topoisomerase 2 α and androgen receptor expression and maximal expression of bcl-2 and E-cadherin. Luminal B subtype showed a high expression of p53, bcl-2 and androgen receptors and moderate expression of topoisomerase 2 α and E-cadherin. High expression of topoisomerase 2 α , low expression of androgen receptors and moderate expression of E- cadherin were registered in triple-negative BC. The maximal expression of topoisomerase 2 α and p53, as well as low expression of bcl-2 and E-cadherin, was observed in Her2+ subtype. Conclusions CD8+ T-lymphocytes dominate in tumor tissue of BC. Besides, tumor tissue of luminal A BC contains more NK-cells than other subtypes. A number of differences in expression of apoptosis-controlling proteins was observed in various molecular subtypes of breast cancer. Luminal A BC was noted for the minimal p53 expression and maximal bcl-2expression. Luminal B subtype had a high p53 and bcl- 2 expression. In Her2+ BC p53 expression was maximal and bcl-2 expression was low. The described differences allow assessing molecular subtypes of breast cancer and predicting the disease course from previously unexplored perspectives.

Ileus secondary to a retroperitoneal malignant melanoma

Retroperitoneal malignant melanomas, either primary or metastatic, are rare. We present our clinical experience concerning one case with ileus secondary to a huge retroperitoneal malignant melanoma. A 77-year-old woman was admitted to the hospital due to progressive abdominal fullness and decreased appetite for 4 months. Plain films showed soft-tissue density in the left lower quadrant of the abdomen, as well as rightward displacement of the intestine. Laboratory data excluded any metabolic or septic causes of ileus. Abdominal computed tomography scan identified a huge retroperitoneal tumor with invasion of the left lower peritoneal space. The mass measured approximately 18.2 × 21.5 cm in the largest section. Immunohistochemical analysis of the tumor biopsies at minilaparotomy showed positive staining of tumor cells for S-100 protein, human melanoma black-45, and vimentin. Thus, a diagnosis of malignant melanoma with peritoneal metastases was established. This case highlights the possibility of a retroperitoneal malignant melanoma exerting a mass effect on the surrounding organs. The authors suggest that malignant melanoma should be taken into consideration as a possible differential diagnosis of retroperitoneal neoplasms.

Global histone modification of H3K27 correlates with the outcomes in patients with metachronous liver metastasis of colorectal cancer

We evaluated the methylation patterns of histone H3 lysine 27 (H3K27), H3 lysine 36 (H3K36) and the expression of H3K27 methylase EZH2 in patients with colorectal carcinomas with metachronous liver metastasis to search for biomarkers identifying these patients. Double 2-mm core tissue microarrays were made from 54 paraffin-embedded samples of primary colorectal adenocarcinomas and corresponding liver metastases and examined using an immunohistochemical analysis of dimethylation and trimethylation in H3K27, H3K36 and EZH2. Positive tumor cell staining for each histone modification (H-score) was used to classify patients into low- and high-staining groups, which were then examined to identify any correlations between the clinicopathological parameters and the clinical outcomes. The H-scores of H3K27me2 were lower in the liver metastases than in the corresponding primary tumors, while the H-scores of H3K36me2 were higher in the liver metastases than in the corresponding primary tumors (P<0.001). H3K27me2 in the primary tumors correlated with tumor size (P=0.016), H3K36me2 in the primary tumors correlated with histological type (P=0.038), and H3K36me3 in the primary tumors correlated with lymph node metastasis (P=0.017). In addition, lower levels of H3K27me2 in the primary tumors correlated with poorer survival rates (P=0.039). The multivariate survival analysis showed that the H3K27me2 status is an independent prognostic factor for colorectal cancer patients (P=0.047). Our findings suggest that the methylation level of H3K27me2 detected with immunohistochemistry may be an independent prognostic factor for metachronous liver metastasis of colorectal carcinomas.

The global histone modification pattern correlates with overall survival in metachronous liver metastasis of colorectal cancer

Post-translational histone modifications are known to be altered in cancer tissues, and differences in the histone modification levels have recently been used to predict the clinical outcome in patients with certain types of cancer. In this study, we evaluated the immunohistochemical staining patterns of histone H3 dimethylation and acetylation in metachronous liver metastasis of colorectal carcinomas and examined its correlation with patient prognosis. Double 2mm core tissue microarrays were made from 54 paraffin-embedded samples of liver metastasis from colorectal adenocarcinoma, and were examined by an immunohistochemical analysis of histone H3 lysine 4 (H3K4) dimethylation, histone, H3 lysine 9 (H3K9) dimethylation and histone H3 lysine 9 (H3K9) acetylation. Positive tumor cell staining for each histone modification was used to classify patients into low- and high-staining groups, which were then examined for correlations with the clinicopathological parameters and clinical outcome. Dimethylation of H3K4 correlated with the tumor histological type (P=0.043), and acetylation of H3K9 correlated with the tumor histological type (P=0.016). In addition, lower levels of H3K4 dimethylation correlated with a poor survival rate (P=0.035). The multivariate survival analysis showed that the H3K4 dimethylation status is an independent prognostic factor for colorectal cancer patients (P=0.011). We suggest that the pattern of histone modification as detected by immunohistochemistry may be an independent prognostic factor for metachronous liver metastasis of colorectal carcinomas.

Metastatic Melanomas in Young Broiler Chickens (Gallus gallus domesticus)

Four young broiler chickens affected by multiple melanotic tumors are described. Grossly, there were multiple tumors composed of melanocytes within the skin, skeletal muscle, and multiple visceral organs. Tumors ranged from flattened macules to masses that extensively replaced viscera. Microscopically, melanocytes were often well pigmented, and while there was moderate nuclear anisokaryosis, mitotic rates were low. Immunohistochemical staining of some melanomas with antibodies to S100 proteins, Melan-A, vimentin, or neuron-specific enolase after bleaching of tumor cells with potassium permanganate revealed lack of immunostaining of tumor cells with antibodies to S100, strong positive staining of tumor cells for neuron-specific enolase, moderate staining with antibodies to vimentin, and faint staining for Melan-A. Only neuron-specific enolase staining was evident in unbleached tumor cells. Attempts to identify exogenous avian leukosis viruses in these tumors were unsuccessful.

Polycythemia and Inappropriate Erythropoietin Concentrations in Two Dogs with Renal T-cell Lymphoma

Two dogs presented with suspected renal disease and polycythemia. Abdominal ultrasound examinations performed on both dogs revealed coalescing masses causing bilateral renomegaly. Serum erythropoietin (EPO) concentrations were physiologically inappropriate. Postmortem examinations revealed renal T-cell lymphoma in both dogs. One of the two dogs also had involvement of the liver and mesentery. EPO-immunohistochemistry on tissue samples demonstrated positive staining in tumor cells and occasional normal renal cells. This report illustrates that paraneoplastic EPO production may induce polycythemia. The pattern of EPO-immunohistochemistry staining suggested that the mechanism of production was due to tumor production of EPO and local hypoxia-induced EPO production from compression of normal renal cells and vasculature.

Abstract P2-09-22: Predictive Significance of the Proportion of ER-or PgR-Positive Tumor Cells in Neoadjuvant Chemotherapy for Operable HER2-Negative Breast Cancer

Abstract Background Estrogen receptor (ER) and progesterone receptor (PgR) status are predictive for clinical and pathological response in neoadjuvant chemotherapy for operable breast cancer. However, it remains unclear how the proportion of ER-or PgR-positive staining tumor cells affect responses to neoadjuvant chemotherapy. The purpose of this study was to examine the correlation of the proportion of ER-or PgR-positive staining tumor cells with the clinical and pathological responses to neoadjuvant chemotherapy for operable HER2-negative breast cancer. Patients and Methods From April 2002 to May 2010, 101 patients received neoadjuvant chemotherapy containing epirubicin and taxane in our clinic. We assessed menopausal status, tumor size, lymph node status, histological grade, ER, PgR, p53 and Ki-67. In this study, we defined the positivity of ER and PgR as more than or equal to 1% of the positive staining tumor cells according to ASCO/CAP announcement in April 2010. Furthermore, patients with ER-positive tumor were divided into two groups at 30% of ER positive staining tumor cells. Results A clinical response was seen in 82% of all patients and a pathological complete response (pCR) was seen in 17%. Of the 101 patients, 60 (59%) had ER-positive tumors and 38 (63%) of the patients had tumors with more than 90% of ER-positive staining cells. Also, 52 (51%) had PgR-positive tumors. Fourteen (34%) of the 41 patients with ER-negative tumors achieved a pCR and 15 (31%) of the 49 patients with PgR-negative tumors achieved a pCR. Patients with more than 30% of ER-or more than 1% of PgR-positive staining tumor cells did not achieve a pCR. There was no significant correlation of pCR with menopausal status, tumor size, grade and Ki-67. In univariate analysis, a pCR was associated with ER status (p=0.0001), PgR status (p=0.0001), and chemotherapy regimens (p=0.014). Multivariate analysis revealed that ER status was a significant factor for pCR, and patients with ER-negative tumors were 20.1 times more likely to achieve a pCR than those with more than or equal to 30% tumors staining tumor cells (p=0.005; 95% confidential interval, 2.5-16.5). Conclusion We demonstrated a predictive significance of the proportion of ER-or PgR-positive tumor cells in neoadjuvant chemotherapy for operable HER2- negative breast cancer. ER-negativity (&amp;lt;1%) is significantly predictive to achieve a pCR in multivariate analysis. Conversely, it seems very possible that patients with more than 30% ER-or more than 1% PgR-positive staining tumor cells do not achieve a pCR. It is important to take the proportion of ER-and PgR-positive staining tumor cells into consideration in the treatment of breast cancer. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-22.

Correlation between chromosome 17 polysomy and estrogen receptor in HER2-positive breast cancer.

e11023 Background: Among HER2 overexpressing breast cancers, chromosome 17 polysomy (C17) may not affect trastuzumab response in several clinical trials. Precise assessment of HER2 is critical in breast cancers and predicts benefit of anti-HER2 therapy. Hormone receptors (HR) and HER2 coexpression in not uncommon in breast cancer, approximately half of HER2 (+) also coexpress HR. Although there are quantitative inverse association between HER2 and both ER and PgR, the biology of triple positive breast cancer and also correlation with C17 are still less known. To analyze the epidemiologic feature of HER2 (+), particularly triple positive in Japanese patients, consecutive operable breast cancer patients during 05-07 treated in single institution were studied. Methods: HER2 status of consecutive 370 patients excluding DCIS and whose specimens were poor quality for study, were analyzed with Pathvysion (FISH). We defined HER2 (+) and C17 as HER2/chromosome 17 ratio > = 2.2 and CEP ratio > = 2.2, respectively. The HR status was evaluated using IHC with the estimated proportion of positive- staining tumor cells according as the Allred score (PS0-5). Results: HER2(+) was found in 19% (71/370) and C17 in 26% (98/370). As regards ER PS2-5 was found in 81% (300/370), particularly PS5 in 68% (251/370). Among 71 HER2(+) cases, PS5 and PS 2-4 were found in 45% (32/71) and 18% (13/71) respectively. Interestingly in HER2(+) cases, there was a trend toward higher FISH ratios with increases in PS score, averages of FISH ratios in PS0-1 (26 cases), PS2-4 (13), and PS5 (32) were 5.9, 6.9, and 7.5 respectively. Although there was no association between the presence of C17 and ER status in HER2(-) cases, ER(+) (PS2-5) was significantly found in higher frequency of 72% (34/47) in HER2(+)C17(-) than 46% (11/24) in HER2(+)C17(+). Conclusions: We found a positive correlation between FISH ratio and proportion of ER positive-staining tumor cells in HER2(+) subset. However, in case of HER2(+), chromosome 17 polysomy drives ER status to negative. No significant financial relationships to disclose.

Cellular Histone Modification Patterns Predict Prognosis and Treatment Response in Resectable Pancreatic Adenocarcinoma: Results From RTOG 9704

PURPOSE Differences in cellular levels of histone modifications have predicted clinical outcome in certain cancers. Here, we studied the prognostic and predictive value of three histone modifications in pancreatic adenocarcinoma. METHODS Tissue microarrays (TMAs) from two pancreatic adenocarcinoma cohorts were examined, including those from a 195-patient cohort from Radiation Therapy Oncology Group trial RTOG 9704, a multicenter, phase III, randomized treatment trial comparing adjuvant gemcitabine with fluorouracil and a 140-patient cohort of patients with stage I or II cancer from University of California, Los Angeles Medical Center. Immunohistochemistry was performed for histone H3 lysine 4 dimethylation (H3K4me2), histone H3 lysine 9 dimethylation (H3K9me2), and histone H3 lysine 18 acetylation (H3K18ac). Positive tumor cell staining for each histone modification was used to classify patients into low- and high-staining groups, which were related to clinicopathologic parameters and clinical outcome measures. Results Low cellular levels of H3K4me2, H3K9me2, or H3K18ac were each significant and independent predictors of poor survival in univariate and multivariate models, and combined low levels of H3K4me2 and/or H3K18ac were the most significant predictor of overall survival (hazard ratio, 2.93; 95% CI, 1.78 to 4.82) in the University of California, Los Angeles cohort. In subgroup analyses, histone levels were predictive of survival specifically for those patients with node-negative cancer or for those patients receiving adjuvant fluorouracil, but not gemcitabine, in RTOG 9704. CONCLUSION Cellular levels of histone modifications define previously unrecognized subsets of patients with pancreatic adenocarcinoma with distinct epigenetic phenotypes and clinical outcomes and represent prognostic and predictive biomarkers that could inform clinical decisions, including the use of fluorouracil chemotherapy.

Evaluation of the frequency of putative prostate cancer stem cells in primary and metastatic prostate cancer

Tumour cells with a stem cell-like phenotype have recently been identified in prostate tumors and it has been suggested that this population may be responsible for the diversity of cell types within tumors and also for the initiation of metastases. These cells carry a number of defined markers: they are cd133 and cd44+ve and express high levels of alpha2beta1 integrin. In this study we have, for the first time, assessed matched primary and bone marrow biopsies from prostate cancer patients for the distribution of cells carrying these and a number of other putative stem cell markers. Eleven matched (primary and bone metastasis) specimens from prostate cancer patients were assessed for the presence of cd133, cd44, alpha2beta1 integrin, CXCR4, c-met, alpha6 integrin, and nestin using immunohistochemistry and stain intensity and distribution scored. In the bone metastases, tumor cells staining positively for cd133 were detected at low frequency in approximately 50% of samples. Staining for nestin was confined to endothelium. Positive staining of tumor cells for the other antigens was present at variable frequency in >70% of metastases with the exception of CXCR4 which was absent from all but 2 specimens. Where positive staining of tumor cells was present in the metastasis, cells staining for each antigen were present in the matched primary with the exception of cd44 which was absent in all but 2/11 matched primary tissues. In established metastases no single or combination of marker expression profiles identify the established metastatic phenotype, although cd44 expression was shown to be more frequent in metastases that in primary cancers.