To investigate the dominant metabolic type of triple-negative breast cancer (TNBC)

To investigate the dominant metabolic type of triple-negative breast cancer (TNBC) and evaluate its clinical implication through analysis of protein expression related to glycolysis glutaminolysis and mitochondrial oxidative phosphorylation. three markers for each phenotype as follows: glycolysis type (Glut-1 CAIX and MCT4) glutaminolysis type (GLS1 GDH and ASCT2) and mitochondrial type (ATP synthase SDHA JTC-801 and SDHB). The Rabbit Polyclonal to Doublecortin (phospho-Ser376). percentages of samples with metabolic phenotypes of tumor and stroma of TNBC were as follows: for tumor mitochondrial type (85.3%) > glutaminolysis type (67.4%) > glycolysis type (63.0%); and for stroma glutaminolysis type (37.2%) > glycolysis type (16.3%) > mitochondrial type (14.0%). The most common metabolic phenotype of TNBC was glycolysis type for basal-like type and non-glycolysis type for non-basal-like type (p=0.047). The correlation between glutaminolysis and mitochondrial type was statistically significant in both tumor and stroma (p<0.001). In conclusion tumor cells of TNBC express glycolysis and mitochondrial metabolism-related proteins. Glycolysis type is the most common phenotype of basal-like type and reversely non-glycolysis type is the most common phenotype of non basal-like type. hybridization (FISH). A cut-off value of 1% or more positively stained nuclei was used to define ER and PR positivity [11]. HER-2 staining was analyzed according to the American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines using the following categories: 0=no immunostaining; 1+=weak incomplete membranous staining less than 10% of tumor cells; 2+=complete membranous staining either uniform or weak in at least 10% of tumor cells; and 3+=uniform intense membranous staining in at least 30% of tumor cells [12]. HER-2 immunostaining was considered positive when strong (3+) membranous staining was observed whereas cases with 0 to 1+ were regarded as negative. The cases showing 2+ HER-2 expression were JTC-801 evaluated for HER-2 amplification by FISH. All the cases were retrospectively reviewed by a breast pathologist (Koo JS) and histological analysis was conducted with hematoxylin and eosin (H&E)-stained slides. The histological grade was assessed using the Nottingham grading system [13]. Clinicopa-thologic parameters evaluated in each case included patient age at initial diagnosis lymph node metastasis tumor recurrence distant metastasis and patient survival. Tissue microarray On H&E-stained slides of tumors a representative area was selected and the corresponding spot was marked on the surface of the paraffin block. Using a biopsy needle the selected area was punched out and a 3-mm tissue core was placed into a 6 × 5 recipient block. JTC-801 The tissue of the invasive tumor was extracted. More than two tissue cores were extracted to minimize the extraction bias. Each tissue core was assigned with a unique tissue microarray location number that was linked to a database containing other clinicopathologic data. JTC-801 Immunohistochemistry The antibodies used for immunohistochemistry in this study are shown in Table 2. All the immunohistochemical assays were conducted with formalin-fixed paraffin-embedded tissue sections. Briefly 5 and/or EGFR-positive) (AR-positive and/or GGT-1-positive) (claudin 3- claudin 4- claudin 7-negative and/or E-cadherin-negative) immune-related type (IL-8-negative and stromal STAT1-positive) (two or JTC-801 more types) and null type (none of these). Metabolic classification of TNBC according to IHC Metabolic phenotypes were classified as follows based on the expression of metabolism-related proteins: (positive for two or more of Glut-1 CAIX and MCT-4) (positive for two or more of GLS1 GDH and ASCT2) and (positive for two or more of ATP synthase SDHA and SDHB). Statistical analysis Data were processed using SPSS for Windows version 12.0 (SPSS Inc. Chicago IL USA). Student’s and Fisher’s exact tests were used to examine any difference in continuous and categorical variables respectively. The limit for statistical significance was set at P=0.05. Kaplan-Meier survival curves and log-rank statistics were employed to evaluate time to tumor metastasis and time to survival. Multivariate regression analysis was performed using.