ABSTRACT
The main findings in the field of breast cancer proteomic research as well as modern strategies, technologies and methods of validation are reviewed. A special attention is focused on validated proteomic biomarkers of breast cancer. The data on proteomic profiling of stroma, tumor microenvironment, involvement of proteins in tumor progression, invasion and metastasis, and mechanisms of action of new generation drugs, are analyzed. The results of proteomic analysis are of high clinical importance and significantly improve tumor molecular profiling, stratification of patients, screening, diagnostics, and therapy of breast cancer.
Subject(s)
Breast Neoplasms/diagnosis , Breast/pathology , Proteins/analysis , Proteomics/methods , Animals , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Biomarkers, Tumor/analysis , Breast/drug effects , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Disease Progression , Female , Humans , Prognosis , Validation Studies as TopicABSTRACT
Using transgenic plants as factories for production of physiologically active human proteins arouses special concern because occasional escape of such transgenes into environment may cause health problems. Creation of plant varieties producing pharmaceutically valuable proteins should be accompanied by development of detection methods suitable for controlling the transgene behavior. Here we describe a multiplex PCR protocol for revealing of two human genes (encoding growth hormone and interferon alpha2b) that have been successfully introduced into plant genomes. The primer pair designed for detection of human growth hormone coding sequence amplifies fragments of different size from the full-length gene in the human genome and the intronless coding sequence usually used for plant transformation. Application of this primer pair may be recommended for ruling out false positive results due to sample contamination with human DNA. Such a control may be useful also in PCR analysis during establishing of transgenic plants carrying genes of human origin.
Subject(s)
DNA Primers/chemistry , Growth Hormone/isolation & purification , Interferon-alpha/isolation & purification , Multiplex Polymerase Chain Reaction/methods , Nicotiana/genetics , Transgenes , Agrobacterium tumefaciens/genetics , Clostridium thermocellum/chemistry , DNA Primers/chemical synthesis , Gene Expression , Glycoside Hydrolases/genetics , Glycoside Hydrolases/metabolism , Growth Hormone/genetics , Growth Hormone/metabolism , Humans , Interferon alpha-2 , Interferon-alpha/genetics , Interferon-alpha/metabolism , Plants, Genetically Modified , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/isolation & purification , Recombinant Fusion Proteins/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Transformation, GeneticABSTRACT
During the last decade interferons are regarded as potent candidates for generation of plant-based edible vaccines because of broad spectrum of antiviral activities and adjuvant properties. Establishment and certification of numerous interferon producing plant systems requests development of fast and efficient multiplex PCR protocol for the transgene detection in GM plants. Here we represent a protocol for simultaneous amplification in one assay of fragments of hIFN alpha 2b gene and two control genes, namely virD1 of Agrobacterium tumefaciens and conservative region of plant actin gene.