flavin-mononucleotide and Breast-Neoplasms

The efficacy of cytoflavin as a supporting drug in the CAF chemotherapy of breast cancer was studied. It was estimated by the tolerability, the peripheric blood index dynamics, the patients general condition (Karnovsky Index) and the frequency of the side effects. For the immediate estimation of the polychemotherapy and cytoflavin effects on the cytoprotection natural system state, the dynamics of the glutathione metabolism and the lipid peroxidation in the erythrocytes was investigated.

Topics: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Breast Neoplasms; Drug Combinations; Female; Flavin Mononucleotide; Humans; Inosine Diphosphate; Middle Aged; Niacinamide; Quality of Life; Succinates

We report a label-free fluorescence lifetime profiling strategy to classify breast cancer cells, MCF10CA1h (malignant), MCF10A (nonmalignant), and MCF10AneoT (premalignant) in different stages of malignancy. Fluorescence Lifetime Imaging Microscopy (FLIM) was used to record the lifetime of autofluorescence of endogenous flavin in MCF10 cells in different stages of malignancy. Predominant differences in lifetimes ascertained by multi-exponential fitting curves can be attributed to the different forms of flavin protein; flavin mononucleotide (FMN), free flavin adenine dinucleotide (FAD), semiquinone, and bound FAD. A lifetime map of the metabolite was derived from the contribution of the lifetime of each metabolite by iterative reconvolution fitting of the Time Correlated Single Photon Counting (TCSPC) decay curves. Lifetime maps were constructed by mapping the average lifetime values pixel by pixel using MATLAB. The FLIM image (150 × 150 pixels) of each cell was extracted, resized and centered into 100 × 100 pixels using the nearest neighbor algorithm. Principal Component Analysis (PCA) in conjunction with Elastic net Analysis (EnA) was then used to classify the different stages of MCF10 cell lines based on average lifetime values. The EnA model provided an excellent classification of the cells at different stages of tumorigenesis yielding 100% accuracy.

Topics: Breast Neoplasms; Cell Line, Tumor; Female; Flavin Mononucleotide; Humans; Microscopy, Fluorescence

Flavoprotein reductases play a key role in electron transfer in many physiological processes. We have isolated a cDNA with strong sequence similarities to cytochrome P-450 reductase and nitric-oxide synthase. The cDNA encodes a protein of 597 amino acid residues with a predicted molecular mass of 67 kDa. Northern blot analysis identified a predicted transcript of 3.0 kilobase pairs as well as a larger transcript at 6.0 kilobase pairs, and the gene was mapped to chromosome 9q34.3 by fluorescence in situ hybridization analysis. The amino acid sequence of the protein contained distinct FMN-, FAD-, and NADPH-binding domains, and in order to establish whether the protein contained these cofactors, the coding sequence was expressed in insect cells and purified. Recombinant protein bound FMN, FAD, and NADPH cofactors and exhibited a UV-visible spectrum with absorbance maxima at 380, 460, and 626 nm. The purified enzyme reduced cytochrome c, with apparent K(m) and k(cat) values of 21 microM and 1.3 s(-1), respectively, and metabolized the one-electron acceptors doxorubicin, menadione, and potassium ferricyanide. Immunoblot analysis of fractionated MCF7 cells with antibodies to recombinant NR1 showed that the enzyme is cytoplasmic and highly expressed in a panel of human cancer cell lines, thus indicating that this novel reductase may play a role in the metabolic activation of bioreductive anticancer drugs and other chemicals activated by one-electron reduction.

Topics: 3T3 Cells; Amino Acid Sequence; Animals; Base Sequence; Binding Sites; Breast Neoplasms; Chromosome Mapping; Chromosomes, Human, Pair 6; Cloning, Molecular; Cytochromes c1; Female; Flavin Mononucleotide; Flavin-Adenine Dinucleotide; FMN Reductase; HeLa Cells; Humans; Kinetics; Mice; Molecular Sequence Data; Molecular Weight; NADH, NADPH Oxidoreductases; NADP; Recombinant Proteins; Sequence Alignment; Sequence Homology, Amino Acid; Spectrophotometry; Tumor Cells, Cultured