A Novel Strategy to Investigate Tissue-Secreted Tumor Microenvironmental Proteins in Serum toward Development of Breast Cancer Early Diagnosis Biomarker Signature.

PURPOSE: The study aims to discover and identify early grade diagnosis markers in tumor microenvironment and investigates their expression in serum. EXPERIMENTAL DESIGN: 2D fluorescence difference gel electrophoresis (2D-DIGE), a gel-based proteomics platform is used for tissue profiling and identifies deregulated proteins by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. Western blot validation for statistically significant six different proteins is performed in an independent cohort of participant serum. RESULTS: Total 67 nonredundant tissue proteins are identified out of 230 protein spots obtained through Marker selection tool of EDA. GELS, DAPLE, IQCC, CATD, A1AT, and HS90B are selected for validation and found to be upregulated in early grade serum samples. Pathway analysis performed through PANTHER and DAVID indicates that apoptosis, CCKR, EGF, FAS, FGF, Wnt, p13 kinase, and p53 signaling pathways are expressed specifically during or early onset of grade I cancer lesions. CONCLUSIONS AND CLINICAL RELEVANCE: GELS, DAPLE, HS90B, A1AT, IQCC, and CATD may be promising potential serum biomarker signature for diagnosis of early grade breast cancer. This panel also contributes to better understanding of molecular mechanisms involved in inceptive stage of breast cancer.

Apoptosis Markers in Breast Cancer Therapy.

Cancer is a disease characterized by a very little apoptosis, ie, genetically programmed cell death. Aberrations in apoptotic pathways are central to tumorigenesis, tumor progression, and overall tumor growth and regression in response to chemotherapy. It is now increasingly accepted that chemotherapeutic drug efficacy is partially related to its ability to induce apoptosis. Apoptosis, therefore, represents not only a vital target in cancer therapy but also a unique biomarker opportunity that has thus far been largely unexploited. In response to therapy, tumor cells undergo apoptosis and release their cellular components in the circulation. As such, these materials may serve as biomarkers to assess response. Apoptosis markers in breast cancer include circulating soluble FasL, granzyme B, and cytochrome c that increase following chemotherapy. Unfortunately, there is a paucity of information in the literature with respect to this approach. As such, large-scale prospective studies are clearly needed to validate this approach and more fully elucidate clinical usefulness.

Quantitative tissue proteomic investigation of invasive ductal carcinoma of breast with luminal B HER2 positive and HER2 enriched subtypes towards potential diagnostic and therapeutic biomarkers.

Worldwide, breast cancer is one of the frequently diagnosed cancers in women with high mortality if not diagnosed at early stage. Although biomarker discoveries through various proteomic approaches have been studied in breast cancer, a limited number of studies have explored the invasive ductal carcinoma with Luminal B HER2 positive (LB) and HER2 enriched (HE) subtypes. The present study employed the complementary quantitative proteomic approaches to find a panel of markers that could discriminate LB and HE subtypes as well as early (ES) and late stages (LS) of these subtypes. A total of 67 and 68 differentially expressed proteins were identified by DIGE for the subtype and stage wise categories, respectively. Multivariate statistical analysis was employed to identify the set of most significant proteins, which could discriminate between these two subtypes and also early and late stages under study. Immunoblotting and MRM based validation in a separate cohort of samples confirmed that panel of biosignatures for LB are APOA1, GELS, HS90B, EF1A1, NHRF1 and PRDX3 and for HE are PRDX1, CATD, CALR, ATPB and CH60. For the diagnosis of early and late stages the potential markers are TPM4, CATD, PRDX3, ANXA3, HSPB1 and CALR, TRFE, GELS, CH60, CAPG, NHRF1, 1433G, GRP78 respectively.

Serum levels of soluble Fas ligand, granzyme B and cytochrome c during adjuvant chemotherapy of breast cancer.

BACKGROUND: Anticancer agents used in chemotherapy for tumors induce apoptosis in malignant cells. Soluble Fas ligand, granzyme B and cytochrome c are key elements in the process of apoptosis. The objective of this preliminary study was to evaluate the changes in the serum concentrations of these parameters in breast cancer patients undergoing adjuvant chemotherapy. MATERIALS AND METHODS: Sixty patients with histopathologically proven breast cancer were included in the present study. The blood samples were taken after surgery before chemotherapy and after 3weeks of administration of the first cycle of chemotherapy. Thirty healthy female controls were selected for comparison. Soluble FasL, granzyme B and cytochrome c were estimated from serum by ELISA. RESULTS: Significantly increased concentrations of soluble FasL, granzyme B and cytochrome c were found in stage II and stage III of breast cancer patients after chemotherapy compared with concentrations before chemotherapy (P<0.0001). A significant positive correlation was found between soluble FasL and cytochrome c as well as between granzyme B and cytochrome c in breast cancer patients after chemotherapy. CONCLUSION: Serum concentrations of apoptotic markers such as soluble FasL, granzyme B and cytochrome c were increased after administration of the first cycle of chemotherapeutic drugs. The measurement of these circulating apoptotic markers may help clinicians in evaluating treatment efficacy in breast cancer.

Ratio Of Serum Asymmetric Dimethyl Arginine (ADMA)/ Nitric Oxide in Coronary Artery Disease patients.

BACKGROUND: Coronary artery disease (CAD) is the leading cause of mortality and morbidity in the world. Current predictions estimate that by the year 2020 cardiovascular diseases, notably atherosclerosis will become the leading global cause of the total disease burden. Atherosclerosis of the Coronary artery causes myocardial infarction and angina pectoris. Endothelial Nitric oxide (NO), released by the intact and healthy endothelium plays a very important role in the maintenance of vascular tone and structure. Decreased NO level leads to endothelial dysfunction is an initial event in the atherosclerosis. Endogenous Asymmetric dimethylarginine (ADMA) is a structural analog of L-arginine, competitively inhibits the enzyme NO synthase and thus decreases the NO level. AIM: To study the ratio of serum ADMA / NO as a marker of severity of CAD. MATERIALS AND METHODS: The study comprises of 60 patients of CAD diagnosed by coronary angiography. We divided them into two Groups according to percentage of atherosclerotic block, Group A (71% and more block, n=30) and Group B (40% - 70% block, n=30). We measured serum ADMA, serum NO and calculated ADMA/ NO ratio. RESULTS were compared with 30 healthy age and sex matched controls. Serum ADMA was determined by reverse phase high performance liquid chromatography. Serum NO was measured by cadmium reduction method. Statistical analysis of data analysis was done using the SPSS (Statistical Package for the Social Science) Version 11 for window. RESULTS: Serum ADMA was correlated positively with the presence and severity of CAD and inversely related with the serum NO levels. Serum ADMA / NO ratio was statistically significant in CAD patients with atherosclerotic block 71% and above (Group A) but ratio was not significant in Group B (block 40% - 70%). CONCLUSION: Serum ADMA/ NO ratio can be the better predictive marker for the severity of the CAD where patient is at the risk of angina pectoris or myocardial infarction due to the extent of coronary atherosclerotic block than individual serum ADMA levels .

Oxidative stress and antioxidant status in patients with alcoholic liver disease.

BACKGROUND: [corrected] Alcoholic liver diseases (ALD) are very common in lower socio-economical strata due to heavy drinking habits and multiple nutritional deficiencies. Ethanol causes liver damage by many mechanisms. The generation of lipid peroxidation by free radicals has been proposed as a mechanism for ethanol induced hepatotoxicity. These free radicals are destroyed by anti-oxidants. Many anti-oxidants are present in the diet, e.g., vitamin E, vitamin C etc. However, poor nutrition or malabsorption leads to deficiency of these vitamins. This may impair the anti-oxidative defense leading to ethanol induced oxidative stress and then to liver damage. METHODS: Oxidative stress and antioxidant defense were assessed in patients with alcoholic liver disease. Serum malondialdehyde (MDA) concentrations were measured as an index of lipid peroxidation, i.e., oxidative stress; and serum vitamins E and C concentrations were measured as an index of antioxidant status. RESULTS: Serum MDA concentrations were increased with the increase in severity of the disease. Concentrations of serum vitamins E and C were decreased in patients with alcoholic liver disease as compared to controls. CONCLUSIONS: Our observations may be due to increased demands of the same or increased utilization.