Tratamiento sistémico para el cáncer de mama HER2 positivo avanzado. Revisión de la evidencia y abordaje terapéutico en el Instituto Nacional de Cancerología - Colombia / Systemic treatment for advanced HER2 positive breast cancer. Review of the evidence and therapeutic approach at the Instituto Nacional de Cancerología - Colombia

El cáncer de mama es la neoplasia más frecuente y de mayor mortalidad en las mujeres en todo el mundo. El receptor 2 del factor de crecimiento epidérmico humano (HER2) se sobreexpresa en aproximadamente el 20% de las pacientes con cáncer de mama y se asocia a mayor riesgo de recidiva tumoral y mortalidad. Antes del desarrollo de los anticuerpos monoclonales dirigidos contra HER2, el cáncer de mama HER2 positivo estaba asociado con un pronóstico desfavorable. El uso de las terapias dirigidas anti HER2 ha mejorado significativamente las tasas de supervivencia global tanto en el escenario adyuvante como en la enfermedad metastásica. En los últimos años han surgido nuevos medicamentos que bloquean esta vía de señalización, lo cual ha permitido establecer varias líneas de tratamiento con terapia anti HER2 en las pacientes con enfermedad metastásica. Por esta razón, las unidades funcionales de Oncología Clínica/Seno y Tejidos Blandos tomaron la decisión de realizar una revisión de la evidencia científica disponible a octubre de 2021, para establecer las recomendaciones en el abordaje terapéutico de las pacientes con cáncer de mama metastásico HER2 positivo en el Instituto Nacional de Cancerología (INC).

Breast cancer is the most common neoplasm and the one with the highest mortality in women worldwide. Human epidermal growth factor receptor 2 (HER2) is overexpressed in approximately 20% of breast cancer patients and is associated with an increased risk of tumor recurrence and mortality. Before the development of monoclonal antibodies directed against HER2, HER2-positive breast cancer was associated with a poor prognosis. The use of anti-HER2 targeted therapies has significantly improved overall survival rates both in the adjuvant setting and in metastatic disease. In recent years, new drugs have emerged that block this signaling pathway, which has made it possible to establish several lines of treatment with anti-HER2 therapy in patients with metastatic disease. For this reason, the clinical oncology/breast and soft tissue functional units made the decision to conduct a review of the available scientific evidence as of October 2021 to establish recommendations for the therapeutic approach to patients with HER2-positive metastatic breast cancer in the National Cancer Institute (INC).

Immunohistochemical Study and Clinicopathologic Correlation of Cox-2 and Her-2 Expression in Colorectal Carcinoma: A 5-Year Retrospective Study

BACKGROUND: Colorectal cancer (CRC) is the fourth most common cancer in Nigeria, and it affects mostly persons in their middle age. In a bid to gain some insight into the molecular characteristics of CRC in our environment, we set out to investigate the expression of COX-2 and HER-2 among Nigerian subjects. OBJECTIVES: To evaluate the expression of COX-2 and HER2 and determine their correlation with clinicopathologic parameters in surgically resected histologically diagnosed cases of colorectal cancer. METHODS: Fifty-three paraffin-embedded tissue blocks of colorectal resections and corresponding patient information were retrieved from the archives of the Anatomic and Molecular Pathology Department of Lagos University Teaching Hospital.A 4-micron slide section was obtained from each specimen and immunohistochemistry for COX-2 and HER-2 expression was performed. RESULTS: Mean age of cases was 53.9years with an almost equal M:F ratio of 1.12:1. Half of the cases were moderately differentiated adenocarcinoma and 17% were high grade tumors.Eighty three percent of the tumours showed positive cytoplasmic COX-2 expression and extremely low membranous HER-2 positivity was observed in 2%. There was no significant correlation between COX-2 expression and age, gender, tumour location, tumour size, depth of invasion or lymph node status.However, COX-2 expression revealed a significant correlation with tumour grade (p= 0.013). CONCLUSION: This study detects a high COX-2 and low HER2 expression in colorectal cancer using immunohistochemistry,suggesting a possible role for COX-2 in CRC pathogenesis.This report should trigger further investigations of both markers vis-à-vis the management of CRC in our environment. WAJM 2022; 39(11): 1134­1140.

L'hyper-expression du HER2 dans le carcinome urothélial de la vessie à propos d'une étude de 30 cas / HER2 hyper-expression in urothelial carcinoma of the bladder in a 30 cases study

Objectif. L'objectif de notre étude est d'évaluer l'expression du HER 2 dans le carcinome urothélial de la vessie sur 30 cas sélectionnés parmi une série de 361 cas colligée au service d'anatomie pathologie du CHU de Batna et de comparer nos résultats aux données de la littérature. Matériels et méthodes. Dans notre étude, nous avons évalué la surexpression de l'HER2 par technique immunohistochimique sur 30 cas, en utilisant les mêmes critères d'interprétation que pour le cancer du sein. Les résultats ont été évalués en utilisant le score d'interprétation de l'American Society of Clinical Oncology (l'ASCO), basé sur le pourcentage de cellules marquées, et l'intensité du marquage dans la perspective d'un traitement par le Trastuzumab (Herceptine). Dans notre étude, seul le score 3 + est considéré comme positif. Résultats et discussion. Nous avons constaté qu'aucun cas de tumeurs à faible potentiel de malignité (TFPM) n'a exprimé le HER2, et que 40 % des carcinomes de haut grade (HG) ont exprimé un marquage intense. Une surexpression de HER2 a été retrouvée dans 23,33% de nos échantillons tumoraux. Ce taux est proche de ceux décrits dans la littérature, allant de 23% à 80% avec d'importants écarts qui pourraient être expliqué par plusieurs hypothèses. Les résultats discordants rapportés dans la littérature nécessitent la standardisation des méthodes des laboratoires. Les différentes études démontrent que les décisions et l'algorithme actuellement utilisés dans les cancers du sein sont également adaptables dans les cancers de la vessie. Conclusion. Malgré la taille réduite de notre échantillon, l'expression de l'HER2 est présente surtout dans les carcinomes de haut grade ce qui concorde avec les données de la littérature des grandes séries. Ces résultats peuvent avoir des implications cliniques sur la prise en charge des tumeurs HER2-positifs localement avancées et/ou métastatiques. Ces patients sont donc des candidats potentiels pour la thérapie ciblée anti HER 2 (Herceptine).

Aim. The aim of the study was to evaluate the expression of HER 2 in urothelial carcinoma of the bladder in 30 cases selected from a series of 361 cases collected at the anatomy pathology department of the CHU of Batna and to compare our results with data from the literature. Method. In our study, we evaluated the overexpression of HER2 immunohistochemically in 30 cases, using the same interpretive criteria as for breast cancer. Results were assessed using the American Society of Clinical Oncology (ASCO) Interpretive Score, based on the percentage of cells labeled, and the intensity of labeling in the perspective of Trastuzumab treatment. (Herceptin). In our study, only the score of 3+ is considered positive. Results and discussion. We found that no cases of low potential malignancy (LPM) tumors expressed HER2, and 40% of high-grade carcinomas (HG) expressed strong staining. Overexpression of HER2 was found in 23.33% of our tumor samples. This rate is close to those described in the literature, ranging from 23% to 80% with large differences that could be explained by several hypotheses. The discordant results reported in the literature require the standardization of laboratory methods. The various studies show that the decisions and the algorithm currently used in breast cancer are also adaptable in bladder cancers. Conclusion. Despite the small size of our sample, the expression of HER2 is present mainly in high-grade carcinomas, which is consistent with data from the literature of large series. These results may have clinical implications for the management of locally advanced and / or metastatic HER2-positive tumors. These patients are therefore potential candidates for targeted anti HER 2 therapy (Herceptin).

Functional and genomic characterization of three novel cell lines derived from a metastatic gallbladder cancer tumor

BACKGROUND: Gallbladder cancer (GBC) is the most common tumor of the biliary tract. The incidence of GBC shows a large geographic variability, being particularly frequent in Native American populations. In Chile, GBC represents the second cause of cancer-related death among women. We describe here the establishment of three novel cell lines derived from the ascitic fluid of a Chilean GBC patient, who presented 46% European, 36% Mapuche, 12% Aymara and 6% African ancestry. RESULTS: After immunocytochemical staining of the primary cell culture, we isolated and comprehensively characterized three independent clones (PUC-GBC1, PUC-GBC2 and PUC-GBC3) by short tandem repeat DNA profiling and RNA sequencing as well as karyotype, doubling time, chemosensitivity, in vitro migration capability and in vivo tumorigenicity assay. Primary culture cells showed high expression of CK7, CK19, CA 19-9, MUC1 and MUC16, and negative expression of mesothelial markers. The three isolated clones displayed an epithelial phenotype and an abnormal structure and number of chromosomes. RNA sequencing confirmed the increased expression of cytokeratin and mucin genes, and also of TP53 and ERBB2 with some differences among the three cells lines, and revealed a novel exonic mutation in NF1. The PUC-GBC3 clone was the most aggressive according to histopathological features and the tumorigenic capacity in NSG mice. CONCLUSIONS: The first cell lines established from a Chilean GBC patient represent a new model for studying GBC in patients of Native American descent.

Hibridación in situ fluorescente (FISH) en el Instituto Nacional de Cancerología (INC) de Colombia. Experiencia de 5 años / Fluorescent in situ hybridization (FISH) in the National Institute of Cancerology (INC) of Colombia. 5 years experience

Resumen Introducción: La hibridación in situ fluorescente (FISH) es una herramienta fundamental en oncopatología para confirmar el diagnóstico de algunas patologías, al igual que determinar el pronóstico y el tratamiento. Objetivo: Describir la experiencia del Instituto Nacional de Cancerología de Colombia (INC) con la técnica de FISH en las diferentes neoplasias hematológicas y tumores sólidos para conocer el comportamiento molecular de nuestra población. Materiales y métodos: Se realizó un estudio descriptivo retrospectivo de todos los resultados de FISH que se han realizado en tumores hematológicos y tumores sólidos en el laboratorio de Genética y Oncología Molecular del INC, entre 2012 y 2016. Resultados: En total se realizaron 1.713 pruebas de FISH, 1.010 (59%) fueron desarrolladas en neoplasias de origen hematolinfoide y 703 (41%) en tumores sólidos, de estos 428 (61%) correspondieron para HER2 de cáncer de seno. En tumores de tejidos blandos fueron evaluadas las sondas MDM2/CDK4, EWSR1, SS18, FUS, CHOP observando positividad en el 10%, el 43%, el 44%, el 20% y el 63%, respectivamente. En cáncer de pulmón se observó positividad en el 12%. Además se realizó estudios para la detección de melanoma y para la detección la codeleción del 1p/19q en gliomas. Discusión: En el INC de Colombia se confirmó la utilidad de la técnica de FISH como complemento en el diagnóstico, el pronóstico y el factor predictivo en el manejo de pacientes con cáncer. Observamos que la prevalencia de algunas pruebas varían de la reportadas en la literatura médica (C-MYC para linfomas, ALK para cáncer de pulmón).

Abstract Introduction: Fluorescent in situ hybridization (FISH) is a fundamental tool in oncopathology to confirm the diagnosis of some pathologies, as well as to determine the prognosis and treatment. Keywords: FISH; Hybridization; Lymphomas; Leukemia; Sarcomas; HER2 Objective: To describe the experience of the FISH in the National Institute of Cancerology of Colombia (INC) in different hematological malignancies and solid tumors to know the molecular behavior of our population. Materials and methods: A retrospective descriptive study was conducted of all the FISH results that have been carried out in the Genetics and Molecular Oncology laboratory of the INC between 2012 and 2016 in hematological tumors and solid tumors. Results: A total of 1713 FISH tests were performed, 1010 (59%) were developed in neoplasms of hematolymphoid origin and 703 (41%) in solid tumors, of these 428 (61%) corresponded to breast cancer (HER2). In soft tissue tumors, MDM2 / CDK4, EWSR1, SS18, FUS, CHOP probes were evaluated, observing positivity in 10%, 43%, 44%, 20% and 63%, respectively. In lung cancer, it has observed positivity in 12%. In addition, studies have been carried out to detect melanoma and to detect the 1p / 19q deletions in gliomas. Discussion: The INC of Colombia confirms the usefulness of the FISH technique as a complement in the diagnosis, prognosis and predictive factor in the management of patients with cancer. We observed that the prevalence of some tests varies from that reported in the medical literature (C-MYC for lymphomas, ALK for lung cancer).

Association between the HER2 Gene Status and the Efficacy of First-line Pemetrexed Combined with Platinum Chemotherapy in Patients with Advanced Lung Adenocarcinoma / 中国肺癌杂志

BACKGROUND@#Human epidermal growth factor receptor 2 (HER2) is one of the driver genes of non-small cell lung cancer (NSCLC). Several studies have shown that the efficacy of pemetrexed in HER2-mutant NSCLC is controversial. The aim of this study is to investigate the efficacy of pemetrexed combined with platinum chemotherapy in patients with HER2-mutant and HER2 wild-type lung adenocarcinoma.@*METHODS@#The clinical data of 106 cases of EGFR, ALK, ROS-1, KRAS, BRAF, RET and MET-negative patients with advanced lung adenocarcinoma patients who diagnosed by histopathology in the First Affiliated Hospital of Zhengzhou University were retrospectively reviewed. The relationships between HER2 gene status, clinical characteristics and response and progression-free survival (PFS) were analyzed.@*RESULTS@#All of the 106 patients' HER2 status were determined. HER2 mutations occurred in 32 cases (30.2%), no mutations in 74 cases (69.8%). HER2 mutations were common in young, non-smoking and female patients. All patients received first-line pemetrexed and platinum-based chemotherapy. The objective response rate (ORR) and disease control rate (DCR) of patients with HER2-mutant lung adenocarcinoma were significantly higher than those without HER2 mutations (40.6% vs 14.9%, χ²=8.464, P=0.004; 93.8% vs 68.9%, χ²=6.327, P=0.012), and the difference was statistically significant. According to univariate analysis, the PFS was significantly associated with the brain metastases, maintenance chemotherapy and HER2 gene status (P0.05). Cox multivariate analysis indicated that HER2 mutation was an independent positive prognostic factor of PFS (P=0.038).@*CONCLUSIONS@#HER2-mutant lung adenocarcinoma patients with first-line pemetrexed combined with platinum chemotherapy have greater clinical benefit than HER2 wild-type patients.

Predictive role of molecular subtypes in response to neoadjuvant chemotherapy in breast cancer patients

Background: Breast cancer is increasingly regarded as a heterogeneous disease which can be classified into distinct molecular subtypes with prognostic significance

Objectives: Retrospective evaluation of the response to neoadjuvant chemotherapy for patients with the major molecular subtypes of breast cancer as classified using immunohistochemical assay and to investigate the patterns of benefit from the neoadjuvant chemotherapy in different molecular subtypes

Materials and methods: ER, PR, HER2 and ki-67 were used to divide102 breast cancer patients treated with neoadjuvant chemotherapy [NCT] into 4 subtypes: luminal A [ER+,PR+,HER2-, and ki-67 14% ; ER+ and/or PR+, HER2+], HER2-overexpression [ER-, PR- and HER2+] and triple-negative [ER-, PR-,and HER2-]

Results: Of the 102 patients analyzed, 9 patients [8.8% of all patients] achieved pCR with 2.6% [2/76] for luminal subgroup, 0.0% [0/8] for HER2-overpression subgroup and 38.9% [7/18] for triple-negative subgroup with a high statistical significant value [p=0.000]

Respuesta al Tratamiento Neoadyuvante en pacientes con Cáncer de Mama en SOLCA-Guayaquil / Response to Neoadjuvant Treatment in patients with Breast Cancer in SOLCA Guayaquil

Introducción: La quimioterapia neoadyuvante es ampliamente aceptada como tratamiento de elección en cáncer de mama localmente avanzado. El objetivo de la presente comunicación corta es conocer la tasa de respuesta completa patológica (pRC) luego de neoadyuvancia, la frecuencia de cirugía conservadora, así como también el porcentaje de la sobrevida. Métodos: Ingresaron al estudio pacientes con cáncer de mama que recibieron tratamiento de quimioterapia neoadyuvante y luego sometidas a cirugía, fueron analizadas retrospectivamente usando historias clínicas desde enero 2009 hasta diciembre 2011 en el Instituto Oncológico Nacional Dr. ¨Juan Tanca Marengo¨ Solca-Guayaquil, se excluyeron pacientes sin suficiente información clínica y aquellas tratadas en otros centros. El procesamiento de datos se realizó mediante un sistema estadístico SPSS v20. Resultados: 1367 pacientes con cáncer de mama fueron diagnosticadas desde el año 2009-2011, se excluyeron aquellas que no reunieron los criterios de inclusión. 200 pacientes evaluables recibieron quimioterapia neoadyuvante y fueron operadas. La edad promedio al diagnóstico fue de 51 años (rango 26-79 años), el tipo histológico ductal Infiltrante fue el más frecuente 185 (92.5 %), Lobular 10 (5 %), Medular 4 (2 %) y metaplásico 1 (0.5 %). El Grado histológico II 143 (71.5 %), Grado histológico III 30 (15 %). En Estadio I: 1 (0.5 %), IIA: 25 (12.5 %), IIB: 42 (21 %), IIIA: 58 (28 %), IIIB: 69 (34.5 %), IIIC: 5 (2.5 %). Conclusión: El cáncer de mama es un grupo heterogéneo, la mayoría de pacientes acuden con enfermedad avanzada (65 %), respuesta patológica completa (16.5 %) luego de neoadyuvancia es muy similar a la obtenida en otros estudios, como es conocido el grupo triple negativo obtuvo los mejores resultados (25 %) y una mejoría en el porcentaje de la sobrevida global en este subgrupo, es importante completar el protocolo de neoadyyuvancia previo a la cirugía para aumentar la tasa de pRC y así como también la cirugía conservadora (10 %) que es el objetivo primario en neoadyuvancia.

Introduction: Neoadjuvant chemotherapy is widely accepted as the treatment of choice in locally advanced breast cancer. The objective of this short communication is to know the pathological complete response rate (pRC) after neoadjuvant, the frequency of conservative surgery, as well as the percentage of survival. Methods: Patients with breast cancer who received neoadjuvant chemotherapy treatment and then underwent surgery, were retrospectively analyzed using clinical histories from January 2009 to December 2011 at the National Cancer Institute "Dr. Juan Tanca Marengo" Solca-Guayaquil, were excluded from the study patients without sufficient clinical information and those treated in other centers. Data processing was performed using a statistical system SPSS v20. Results: 1367 patients with breast cancer were diagnosed from the year 2009-2011, those who did not meet the inclusion criteria were excluded. 200 evaluable patients received neoadjuvant chemotherapy and were operated on. The average age at diagnosis was 51 years (range 26-79 years), the intracranial ductal histological type was the most frequent 185 (92.5 %), Lobular 10 (5 %), Medular 4 (2 %) and metaplastic 1 (0.5 %). The histological grade II 143 (71.5 %), histological grade III 30 (15 %). In Stage I: 1 (0.5 %), IIA: 25 (12.5 %), IIB: 42 (21 %), IIIA: 58 (28 %), IIIB: 69 (34.5 %), IIIC: 5 (2.5 %). Conclusion: Breast cancer is a heterogeneous group, most patients come with advanced disease (65 %), complete pathological response (16.5 %) after neoadjuvant is very similar to that obtained in other studies, as is known the triple negative group obtained the best results (25 %) and an improvement in the percentage of overall survival in this subgroup, it is important to complete the neoadjuvant protocol prior to surgery to increase the pRC rate as well as conservative surgery (10 %) it is the primary objective in neoadjuvant.

Efectos nucleares del receptor Tirosina Quinasa ErbB-2 en cáncer de mama / NUCLEAR EFFECTS OF THE ErbB-2 RECEPTOR TYROSINE KINASE IN BREAST CANCER

Aproximadamente 15-20% de los cánceres de mama (CM) presentan sobre- expresión en la membrana citoplasmática de ErbB-2 (MErbB-2), un miembro de la familia ErbBs de receptores con actividad de tirosina quinasa, o bien presentan amplificación del gen. Antes del desarrollo de terapias dirigidas contra el MErbB-2, este subtipo de CM, denominado ErbB-2-positivo, estaba asociado con un aumento en el potencial metastásico del tumor y un mal pronóstico. Estas terapias han aumentado significativamente la sobrevida global y el porcentaje de enfermos curados. Sin embargo, la resistencia a las terapias disponibles actualmente es todavía un importante problema en la clínica. Actuando por su mecanismo clásico, el MErbB-2 activa cascadas de señalización que transducen sus efectos en el cáncer de mama. La presencia del ErbB-2 en el núcleo fue descubierta hace más de veinte años. Evidencias experimentales proporcionadas por varios grupos de investigación, incluyendo el nuestro, revelaron una función no canónica del ErbB-2 en el núcleo celular donde actúa como un regulador de transcripción. Nuestros hallazgos demostraron que el ErbB-2 nuclear estimula el crecimiento del CM, el desarrollo de metástasis y la resistencia a las terapias utilizadas actualmente

Membrane overexpression of ErbB-2 (MErbB-2), a member of the ErbBs family of receptor tyrosine kinases, or ErbB-2 gene amplification, occurs in 15-20% of breast cancers (BC). Until the development of MErbB-2-targeted therapies, this BC subtype, called ErbB-2-positive, was associated with increased metastatic potential and poor prognosis. Although the overall survival and cure rates have improved significantly with such therapies, resistance to available drugs is still a major clinical issue. In its classical mechanism, MErbB-2 activates downstream signal cascades, which transduce its effects in BC. The fact that ErbB-2 is also present at the nucleus of BC cells was discovered over twenty years ago. Also, compelling evidence revealed a non-canonical function of nuclear ErbB-2 as a transcriptional regulator. Since deeper understanding of nuclear ErbB-2 actions would be critical to disclose its role as a biomarker and a target of therapy in BC, we will here review its function in BC, focusing on its role in growth, metastatic spreading, and response to currently available MErbB-2 positive BC therapies.

Modified allele-specific PCR improves HER2 Ile655Val detection by reducing genotyping errors

Background: A reliable method to detect gene polymorphisms must be established to eliminate genotyping errors due to false PCR amplification. In the previous study, we have developed AS-PCR (Allele Specific-Polymerase Chain Reaction) to detect HER2 Ile655Val gene polymorphism with good specificity and sensitivity, yet it produces some errors. This study is aimed to eliminate the source of genotyping errors mainly by betaine treatment and PCR program modification. Methods: Genotyping errors produced by AS-PCR was qualitatively and quantitatively evaluated using two genomic DNA that each contained AA genotype and GG genotype of HER2 Gene. Betaine treatment or PCR program modification was tested to eliminate the occurrence of genotyping errors during AS-PCR amplification. Results: The types of genotyping errors exhibited by HER2 Ile655Val AS-PCR are diverse, ranging from LDO (Locus Drop Out), preferential amplification to ADO (Allele Drop Out). The rate of genotyping errors was from 10% to 50% depending on the amount and ratio of DNA template and the annealing temperature of PCR. In the mixed DNA template model, the betaine treatment has shown to reduce ADO only in preferentially amplified GG genotype amplicon. Alternatively, reducing the template of the heterozygous DNA by half ( -0.5 ng of DNA template) for such case has effectively recovered the AS-PCR from ADO. Furthermore, increasing the denaturation temperature to 96 °C with an annealing time of 40 s at the first 10 cycles of AS-PCR has succeeded in eliminating severe preferential amplification of AA genotype amplicon by preventing the DNA template with GG genotype from forming into a G-quadruplex structure. The guideline offered in this study has been successfully applied for clinical samples of breast cancer that show preferential amplification. Conclusion: Betaine and the modifying AS-PCR program can reduce significantly genotyping errors making AS-PCR for HER2 Ile655Val detection more reliable to be used as a molecular tool for genotyping purpose (AU)

HER2 Status and Its Heterogeneity in Gastric Carcinoma of Vietnamese Patient

BACKGROUND: Human epidermal growth factor receptor 2 (HER2) is related to the pathogenesis and poor outcome of numerous types of carcinomas, including gastric carcinoma. Gastric cancer patients with HER2 positivity have become potential candidates for targeted therapy with trastuzumab. METHODS: We investigated 208 gastric cancer specimens using immunohistochemistry (IHC), fluorescence in situ hybridization and dual in situ hybridization (ISH). We also investigated the concordance between IHC and ISH. The correlation between HER2 status and various clinicopathological findings was also investigated. RESULTS: In total, 15.9% (33/208) and 24.5% (51/208) of gastric cancers showed HER2 gene amplification and protein overexpression, respectively. A high level of concordance between ISH and IHC analyses (91.3%, κ = 0.76) was found. A significant correlation between HER2 status and intestinal-type (p < .05) and differentiated carcinomas (p < .05) was also noted. The HER2 heterogeneity was high in gastric cancers; we found 68.8% phenotypic heterogeneity and 57.6% genotypic heterogeneity. Heterogeneity in HER2 protein expression and gene amplification showed a close association with diffuse histologic type and IHC 2+. CONCLUSIONS: HER2 protein overexpression and gene amplification were detected in 24.5% and 15.9% of gastric cancer specimens, respectively. Intestinal-type showed a higher level of HER2 protein overexpression and gene amplification than diffuse type. HER2 status also showed a significant relationship with well- and moderately-differentiated carcinomas. The ratio of phenotypic and genotypic heterogeneity of HER2 was high in gastric carcinomas and was associated with HER2 IHC 2+ and diffuse histologic type.

MicroRNA-222 Expression as a Predictive Marker for Tumor Progression in Hormone Receptor-Positive Breast Cancer / 한국유방암학회지

PURPOSE: The microRNA-221/222 (miR-221/222) gene cluster has been reported to be associated with the promotion of epithelial-mesenchymal transition (EMT), downregulation of estrogen receptor-α, and tamoxifen resistance in breast cancer. We studied the expression of miR-222 in human breast cancer samples to analyze its relationship with clinicopathologic features of the tumor, including estrogen receptor status, expression of EMT markers, and clinical outcomes. METHODS: Quantitative real-time polymerase chain reaction was performed to detect the expression of miR-222 in 197 invasive breast cancers. Expression of EMT markers (vimentin, smooth muscle actin, osteonectin, N-cadherin, and E-cadherin) was evaluated using immunohistochemistry. RESULTS: High miR-222 levels were associated with high T stage, high histologic grade, high Ki-67 proliferation index, and HER2 gene amplification. Its expression was significantly higher in the luminal B and human epidermal growth factor receptor 2-positive (HER2+) subtypes than in the luminal A and triple-negative subtypes. In the hormone receptor-positive subgroup, there was a significant negative correlation between miR-222 and estrogen receptor expression, and miR-222 expression was associated with EMT marker expression. In the group as a whole, high miR-222 expression was not associated with clinical outcome. However, subgroup analyses by hormone receptor status revealed that high miR-222 expression was a poor prognostic factor in the hormone receptor-positive subgroup, but not in the hormone receptor-negative subgroup. CONCLUSION: This study showed that miR-222 is associated with down-regulation of the estrogen receptor, EMT, and tumor progression in hormone receptor-positive breast cancer, indicating that miR-222 might be associated with endocrine therapy resistance and poor clinical outcome in hormone receptor-positive breast cancer.

Human epidermal growth factor receptor expression in colorectal cancer and its relationship with clinicopathological characteristics

Background: Some recent studies reported human epidermal growth factor receptor [HER-2/neu] as a marker that can be used in immunological studies of colorectal carcinoma for predicting the prognosis and the treatment. Therefore, we aimed to investigate the frequency of HER-2 expression in patients with colorectal cancer, and explore the relationship between clinicopathological prognostic factors and its expression based on immunohistochemical analysis

Methods: This study included 50 patients with a histologically proven diagnosis of colorectal carcinoma who received surgery at Imam Khomeini Hospital affiliated to Mazandaran University of Medical Sciences. First, HER-2/neu protein expressions were detected by immunohistochemistry and then the data extracted from recorded files

Results: The median age of the patients was 60.2 +/- 13.9 years [range: 25-93 years]. There was no significant relationship between size of tumor, age, sex, lymph node metastases, distant metastasis, differentiation, and stage of the disease with positive expression of HER-2 in this study

Conclusion: No significant relationship between expression of HER-2 and clinicopathological prognostic factors was found in our study. Further comprehensive and prospective trial with standard method to evaluate the role of HER-2 expression among patients with colorectal cancer is needed

Comparing the effect of silybin and silybin advanced[TM] on viability and HER2 expression on the human breast cancer SKBR3 cell line by no serum starvation

The polyphenol silybin has anti-oxidant and anti-cancer properties. The poor bioavailability of some polyphenols [flavonoids, and terpenoids] can be improved by binding them to phosphatidylcholine [phytosome technology]. Many studies have focused on the most common phytosome, silybin-phosphatidylcholine, particularly for its hepatoprotective effects. However, in recent years, studies have also been conducted to determine its anti-cancer effect. Considering that the serum starvation should not be used for studies that are not focused on cell cycle arrest, we studied the effect of silybin-phosphatidylcholine from Silybin Advanced[TM] in 1:2 ratio [one part silybin bound to two parts phosphatidylcholine] on HER2 gene expression on the SKBR3 breast cancer cell line which were cultured in complete medium [not serum deprivation]. The results were compared with our previous study of silybin on HER2 expression on SKBR3 cells. An MTT test was used to determine concentrations for cell treatment, and the gene expression was defined by real-time RT-PCR. Outcomes showed significant concentration- and time-dependent cell growth inhibitory effects of silybin, and silybin-phosphatidylcholine and HER2 down regulation on SKBR3 cells. Silybin-phosphatidylcholine concentrations had a much larger inhibitory and HER2 down regulate effect on cell growth than the same silybin concentrations on SKBR3 cells

Detecting the spectrum of multigene mutations in non-small cell lung cancer by Snapshot assay / 癌症

As molecular targets continue to be identified and more targeted inhibitors are developed for personalized treatment of non-small cell lung cancer (NSCLC), multigene mutation determination will be needed for routine oncology practice and for clinical trials. In this study, we evaluated the sensitivity and specificity of multigene mutation testing by using the Snapshot assay in NSCLC. We retrospectively reviewed a cohort of 110 consecutive NSCLC specimens for which epidermal growth factor receptor (EGFR) mutation testing was performed between November 2011 and December 2011 using Sanger sequencing. Using the Snapshot assay, mutation statuses were detected for EGFR, Kirsten rate sarcoma viral oncogene homolog (KRAS), phosphoinositide-3-kinase catalytic alpha polypeptide (PIK3CA), v-Raf murine sarcoma viral oncogene homolog B1 (BRAF), v-ras neuroblastoma viral oncogene homolog (NRAS), dual specificity mitogen activated protein kinase kinase 1 (MEK1), phosphatase and tensin homolog (PTEN), and human epidermal growth factor receptor 2 (HER2) in patient specimens and cell line DNA. Snapshot data were compared to Sanger sequencing data. Of the 110 samples, 51 (46.4%) harbored at least one mutation. The mutation frequency in adenocarcinoma specimens was 55.6%, and the frequencies of EGFR, KRAS, PIK3CA, PTEN, and MEK1 mutations were 35.5%, 9.1%, 3.6%, 0.9%, and 0.9%, respectively. No mutation was found in the HER2, NRAS, or BRAF genes. Three of the 51 mutant samples harbored double mutations: two PIK3CA mutations coexisted with KRAS or EGFR mutations, and another KRAS mutation coexisted with a PTEN mutation. Among the 110 samples, 47 were surgical specimens, 60 were biopsy specimens, and 3 were cytological specimens; the corresponding mutation frequencies were 51.1%, 41.7%, and 66.7%, respectively (P = 0.532). Compared to Sanger sequencing, Snapshot specificity was 98.4% and sensitivity was 100% (positive predictive value, 97.9%; negative predictive value, 100%). The Snapshot assay is a sensitive and easily customized assay for multigene mutation testing in clinical practice.

Assessment of HER2 gene amplification in breast cancer: a comparison of dual-color in-situ hybridization and fluorescence in-situ hybridization / 中华病理学杂志

<p><b>OBJECTIVE</b>To compare dual-color in-situ hybridization (DISH) with fluorescence in situ hybridization(FISH) in evaluating the human HER2 gene status in invasive breast cancer.</p><p><b>METHODS</b>HER2 gene status in 110 cases of breast invasive ductal carcinomas with a 2+ score on immunohistochemistry (IHC) was investigated by FISH and DISH. The 2007 and 2013 ASCO (American Society of Clinical Oncology)/CAP (College of American Pathologists) HER2 guideline were used respectively to evaluate the agreement between these two techniques.</p><p><b>RESULTS</b>(1) Using the 2007 ASCO/CAP guideline, the overall concordance between FISH and DISH was 97.3% (107/110). Fifty of 51 samples with amplification by FISH were also detected by DISH and the remaining one sample was equivocal.Eight of 10 equivocal samples by FISH were equivocal by DISH and the remaining two samples were negative. Forty-nine samples with no amplification by FISH were all negative by DISH. The concordance was 98.0%, 8/10 and 100.0% respectively for the FISH positive, equivocal and negative groups. (2) Using the 2013 ASCO/CAP guideline, the overall concordance between FISH and DISH was 90.0% (99/110). Fifty-three of 55 samples with amplification by FISH were also detected by DISH and the remaining two were equivocal and negative respectively. Two of 12 equivocal samples by FISH were equivocal by DISH and the remaining ten samples were negative in 7 cases and equivocal in 3 cases. Forty-three samples with no amplification by FISH were all negative by DISH. The concordance was 96.4%, 3/12 and 100.0% respectively for the FISH positive, equivocal and negative groups. Pearson correlation analysis indicated that the HER2 status detected by FISH and DISH were significantly correlated with each other (R=0.584, P<0.01).</p><p><b>CONCLUSIONS</b>There is a high concordance between DISH and FISH for assessing the HER2 gene status in invasive breast cancer. DISH is a new option for assessing HER2 gene status of breast cancer in clinical practice. The clinical significance of the discordance between DISH and FISH in equivocal cases warrants further study.</p>

Dual-color silver-enhanced in-situ hybridization for determination of HER2 gene amplification in gastric carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the concordance of dual-color silver enhanced in-situ hybridization (DSISH) and immunohistochemistry (IHC) in the detection of HER2 gene amplification and expression and to evaluate the values of DSISH in detecting HER2 gene status in gastric carcinoma.</p><p><b>METHODS</b>By using automated DSISH and IHC, HER2 gene status was detected in 230 cases of gastric cancer.</p><p><b>RESULTS</b>Among the 230 cases of gastric carcinoma tested by DSISH, 43 cases were positive and 187 cases were negative; HER2 gene amplification rate was 18.7% (43/230). The expression of HER2 protein was negative, weakly, moderately and strongly positive in 115, 69, 15 and 31 cases, respectively, by IHC. HER2 protein positive rate was 13.5% (31/230). Of the 43 HER2 gene amplification cases by DSISH, 2, 10, 2 and 29 cases were negative, weakly, moderately and strongly positive by IHC; Of the 187 HER2 negative cases by DSISH, 113, 59, 13 and 2 cases were negative, weakly, moderately and strongly positive by IHC, respectively. The overall concordance of HER2 status in the investigation between IHC and DSIDH was 93.5% (201/215), with a high consistency (Kappa coefficient 0.767, P < 0.01).</p><p><b>CONCLUSIONS</b>DSISH can be applied to detect the HER2 gene status in gastric cancer and it also has a high consistency with the result of IHC. In addition, due to frequent heterogeneous expression of HER2, cases with moderate HER2 protein expression may need further assessment by DSISH.</p>

Immunohistochemical expression of Her2/Neu receptor in human colorectal carcinoma [a clinicopathological study]

Colorectal adenocarcinoma is the most common type of gastrointestinal cancers in Iraq, and according to the Iraqi cancer registry [ICR] reports, the incidence of colorectal carcinoma was 2.99% of whole body malignancies [ICR 2010]. In males, it's the 5[th] common cancer while in females it's the 4[th] most common cancer. Her2/neu is an important oncogene in breast cancer, but its prevalence and significance in colorectal carcinoma have been documented. To determine the frequency and the pattern of Her2/neu expression in colorectal adenocarcinoma by immunohistochemical technique and to correlate this expression with different clinicopathological parameters. Twenty five cases of colorectal adenocarcinoma were studied, these cases were diagnosed in private laboratories in Baghdad/Iraq from November 2011 to march 2013. Clinicopathological parameters such as age, gender, pathological diagnosis, including the tumor site, size, lymph nodes status, grade and stage of tumor were taken from patients file. Sections of 4 µm stained by H and E stain and immunohistochemical stained for Her2/neu. Using infiltrative ductal carcinoma of the breast as control positive, evaluation of Her2/neu expression by immunohistochemistry in all cases was performed. Fourteen [56%] of the cases were males, 11 [44%] case were females, with age distribution ranging from [24-89] years, with a mean age of 56.5 years. Tumor size ranges between 2.5-10 cm, with mean of 6.25 cm. Seven [28%] cases were localized in the cecum, 5 [20%] from each rectum, sigmoid and left colon, respectively and 3 [12%] involving more than one segment of the colon. Histologically the tumor grade ranges from moderately differentiated in 23 [92%] cases and poorly differentiated in 2 [8%] cases. Regarding pathological staging [TNM system], 5 [20%] were stage T2, 17 [68%] were stage T3 and 3 [12%] were T4. Lymph node involvement found in 10[40%] of the cases and distant metastasis was found in 2 [12%] cases. Her2/neu expression was present in 4[16%] cases of 25 colorectal adenocarcinoma; there was no correlation with age, sex, histopathological grade, location, lymph nodes status and tumor invasion. Concerning data exists about the prevalence of her2/new expression in colorectal adenocarcinoma, there was no significant correlation between her2/neu over expression and tumor size, grade, localization of the primary tumor, lymph nodes status and depth of invasion

Pattern of human epidermal growth factor receptor 2 [HER2] expression in invasive breast carcinoma, immunohistochemical - study on patients in Upper Egypt

Breast cancer is a major public health problem throughout the world. It accounts for 38% of all new cancer cases among women living in Egypt. One of the most important prognostic and determinant factor of the line of its treatment is the human epidermal growth factor receptor 2 [HER2], it is associated with the more aggressive phenotype. Attention has been focused on the expression of HER2 receptor proteins in breast cancer cells especially its membranous domain, it resulted in variable results concerning its percentage of expression as well as its geographic distribution. So there is a need to study HER2 types of expression in breast cancer patients in our location as well as its correlation with the clinicopathological parameters. HER2 expression in 336 retrospective breast cancer specimens was examined immunohistochemically using tissue microarray. Expression was scored into 0, 1, 2 and 3 degrees and was correlated with clinicopathological criteria. HER2 expression in our specimens showed both membranous and cytoplasmic staining patterns. 18.6% of specimens showed membranous immunoreactivity and 74.1% specimens showed cytoplasmic staining pattern. Significant statistical association was found between cytoplasmic staining of HER2 and tumors of low grade, ER positivity [p<0.001, 0.001, 0.008] respectively. There was statistical significance difference between high membranous expression of HER2 and ER negativity [p=0.038], but our results didn't find significant difference with tumor size, lymphvascular invasion or lymph node metastasis. The frequency of high membranous expression of HER2 in our specimens is 18.6% and inversely correlated with ER positive tumors. This group of patients should be subjected to specific treatment with Trastuzumab, to improve their survival. Surprisingly cutoplasmic expression detected in most of our patients with frequency of 74.1% with positive relationship to low tumor grade and hormone receptor positive tumors. Since this group of patients may be resistant to trastuzumab and need specific treatment with tyrosine kinase drug inhibitors, this observation is going to be discussed and need to be followed up in the future

Quantitative analysis of ErbB1 and ErbB2 genes amplification by a high performance liquid chromatography

Genes for human epidermal growth factor receptors B1 [ErbB1] and B2 [ErbB2] were amplified in breast and ovarian cancers. Both of them were associated with aggressive disease and worse prognosis. The ErbB1 or ErbB2 status of a tumor may provide an indication of the response to ErbB1 and ErbB2 -targeted therapies. For accurate and rapid assessment of amplification of ErbB1 and ErbB2 oncogenes, a High Performance Liquid Chromatography [HPLC] method was developed in this study. DNA was extracted from 30 primary breast tumors and 20 blood samples of healthy donors. ErbB1 and ErbB2 genes along with a reference gene were co-amplificated by Polymerase Chain Reaction [PCR]. The PCR products were separated and quantified using an anion- exchange column within 30 min and in a single step. Optimum resolution was obtained when a sodium chloride gradient and a column temperature of 35°C were used. The results of HPLC analysis of ErbB1 and ErbB2 PCR products were compared with real time PCR method as a gold standard test for 7 tumor samples. The proposed HPLC method was confirmed by real time PCR method. Twenty two and ten of the specimens in our breast cancer cohort showed more than a two-fold amplification of ErbB2 and ErbB1 oncogenes, respectively. Our results were confirmed by real time PCR and showed that HPLC method is a specific, cheap and clinically applicable analytical approach for assessment of ErbB1 and ErbB2 statuses in breast tumors