ABSTRACT
Antitumor efficacy of the combined suicide gene therapy and radiotherapy was studied on the model of CT26 murine colon adenocarcinoma. CMV-FCU1-IRES-mGM-CSF-pGL3 construct with PEG-PEI-TAT (FCU1-mGM/5-FC) block copolymer as a vector was used for intratumoral administration. Tumors were irradiated with a single 5 Gy dose. The efficacy was evaluated according to the grade of tumor growth inhibition (T/C) and lifespan of the animals. Pronounced antitumor activity of the combined use of FCU1-mGM/5-FC system with radiotherapy on the background of prolonged lifespan and the synergism of the applied methods was revealed.
Subject(s)
Adenocarcinoma/therapy , Colonic Neoplasms/therapy , Genes, Transgenic, Suicide , Genetic Therapy/methods , Adenocarcinoma/pathology , Animals , Antimetabolites, Antineoplastic/administration & dosage , Cell Line, Tumor , Colonic Neoplasms/pathology , Combined Modality Therapy/methods , Cytomegalovirus/genetics , Flucytosine/administration & dosage , Fluorouracil/administration & dosage , Genetic Vectors , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Mice, Inbred BALB C , Neoplasm Grading , Neoplasm Transplantation , Treatment Outcome , Tumor BurdenABSTRACT
The embryonic development and carcinogenesis are controlled by many transcription factors. The regulatory systems involved in embryogenesis of an organ are also involved in the tumor development in the same organ. FOX family proteins are transcription factors, which play a key role in these processes. The pioneering factors of the FOXA subfamily act at the very early stages of the embryonic development by interacting with condensed chromatin and thereby enabling the expression of the formerly silent important transcription factors. The role of these factors in tumor development is currently not fully elucidated, although recent studies indicate the important contribution of the FOXA subfamily proteins at the early stages of carcinogenesis. This review is restricted to the role of the FOXA factors in embryogenesis of the pancreas and their significance in the development of the pancreatic ductal adenocarcinoma.
Subject(s)
Cell Transformation, Neoplastic , Embryo, Mammalian , Embryonic Development , Hepatocyte Nuclear Factors , Neoplasm Proteins , Pancreatic Neoplasms , Animals , Cell Transformation, Neoplastic/genetics , Cell Transformation, Neoplastic/metabolism , Cell Transformation, Neoplastic/pathology , Embryo, Mammalian/metabolism , Embryo, Mammalian/pathology , Hepatocyte Nuclear Factors/genetics , Hepatocyte Nuclear Factors/metabolism , Humans , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/metabolism , Pancreatic Neoplasms/pathologyABSTRACT
The review discusses the causes of multiple failures in cancer treatment, which might primarily result from the excessive variability of cancer genomes. They are capable of changing their spatial and temporal architecture during tumor development. The key reasons of irreproducibility of biomedical data and the presumable means for improvement of therapeutic results aiming at targeting the most stable tumor traits are suggested.
Subject(s)
Molecular Biology , Neoplasms/genetics , Animals , Humans , Neoplasms/pathology , Neoplasms/therapy , Reproducibility of ResultsABSTRACT
Great successes in identification and deciphering of mechanisms of the adult stem cells regulation have given rise to the idea that stem cells can also function in tumors as central elements of their development, starting from the initial stage and continuing until metastasis. Such cells were called cancer stem cells (CSCs). Over the course of intense discussion, the CSCs hypothesis gradually began to be perceived as an obvious fact. Recently, the existence of CSCs has been indeed confirmed in a number of works. However, when are CSCs universal prerequisites of tumors and to what extent their role is essential for tumor evolution remains an issue far from resolved. Likewise, the problem of potential use of CSCs as therapeutic targets remains unsolved. The present review attempts to analyze the issue of cancer stem cells and the potential of targeting them in tumor therapy.
ABSTRACT
Core promoters with adjacent regions of the human genes CDC6, POLD1, CKS1B, MCM2, and PLK1 were cloned into a pGL3 vector in front of the Photinus pyrails gene Luc in order to study the tumor specificity of the promoters. The cloned promoters were compared in their ability to direct luciferase expression in different human cancer cells and in normal fibroblasts. The cancer-specific promoter BIRC5 and non-specific CMV immediately early gene promoter were used for comparison. All cloned promoters were shown to be substantially more active in cancer cells than in fibroblasts, while the PLK1 promoter was the most cancer-specific and promising one. The specificity of the promoters to cancer cells descended in the series PLK1, CKS1B, POLD1, MCM2, and CDC6. The bidirectional activity of the cloned CKS1B promoter was demonstrated. It apparently directs the expression of the SHC1 gene, which is located in a "head-to-head" position to the CKS1B gene in the human genome. This feature should be taken into account in future use of the CKS1B promoter. The cloned promoters may be used in artificial genetic constructions for cancer gene therapy.
ABSTRACT
Head-to-head genes with a short distance between their transcription start sites may constitute up to 10% of all genes in the genomes of various species. It was hypothesized that this intergenic space may represent bidirectional promoters which are able to initiate transcription of both genes, but the true bidirectionality was proved only for a few of them. We present experimental evidence that, according to several criteria, a 269 bp region located between the PSENEN and U2AF1L4 human genes is a genuine bidirectional promoter regulating a concerted divergent transcription of these genes. Concerted transcription of PSENEN and U2AF1L4 can be necessary for regulation of T-cell activity.
Subject(s)
Amyloid Precursor Protein Secretases/genetics , Gene Expression Regulation , Membrane Proteins/genetics , Nuclear Proteins/genetics , Promoter Regions, Genetic , Ribonucleoproteins/genetics , Amyloid Precursor Protein Secretases/metabolism , Cell Line , Cloning, Molecular , Consensus Sequence , Humans , Membrane Proteins/metabolism , Nuclear Proteins/chemistry , Nuclear Proteins/metabolism , Nucleotide Motifs , Organ Specificity/genetics , Protein Subunits/chemistry , Protein Subunits/genetics , Protein Subunits/metabolism , Ribonucleoproteins/chemistry , Ribonucleoproteins/metabolism , Splicing Factor U2AF , Transcription, GeneticABSTRACT
Here we directly compared gene expression profiles in human esophageal squamous cell carcinomas and in human fetal esophagus development. We used the suppression subtractive hybridization technique to subtract cDNAs prepared from tumor and normal human esophageal samples. cDNA sequencing and reverse transcription polymerase chain reaction (RT-PCR) analysis of RNAs from human tumor and the normal esophagus revealed 10 differentially transcribed genes: CSTA, CRNN, CEACAM1, MAL, EMP1, ECRG2, and SPRR downregulated, and PLAUR, SFRP4, and secreted protein that is acidic and rich in cysteine upregulated in tumor tissue as compared with surrounding normal tissue. In turn, genes up- and downregulated in tumor tissue were down- and upregulated, respectively, during development from the fetal to adult esophagus. Thus, we demonstrated that, as reported for other tumors, gene transcriptional activation and/or suppression events in esophageal tumor progression were opposite to those observed during development from the fetal to adult esophagus. This tumor 'embryonization' supports the idea that stem or progenitor cells are implicated in esophageal cancer emergence.
Subject(s)
Carcinoma, Squamous Cell/genetics , Esophageal Neoplasms/genetics , Esophagus/embryology , Gene Expression Regulation, Neoplastic/physiology , Adult , Aged , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Case-Control Studies , Cohort Studies , Esophageal Neoplasms/metabolism , Esophageal Neoplasms/pathology , Esophagus/metabolism , Esophagus/pathology , Female , Gene Expression Profiling , Humans , Male , Middle Aged , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The sequences of a 1.5 kb long stretch of the 5' flanking region of the gene for the alpha 3 isoform of the catalytic subunit of human Na(+)-K+ ATPase (located on chromosome 19) and of more than a 2 kb stretch of the 5' flanking region of the gene for the alpha 2 isoform (located on chromosome 1) have been determined. Transcription start sites for the gene for the alpha 3 isoform have been mapped at positions -152 and -155 relative to the translation initiation codon by primer extension analysis and S1-nuclease mapping of mRNA from human brain. The 5' flanking region of the gene for isoform alpha 3 contains a CCAAT box on the noncoding chain and six putative Sp1 binding sites. Absence of a conventional TATA box and a high GC content are other features of the region. The 5' upstream region of the gene for the alpha 2 isoform contains potential TATA and CCAAT boxes and one potential Sp1 binding site. Upstream of the putative TATA box there is an octanucleotide repeat, GGGGGAGA, which is also found in several eukaryotic genes in analogous positions. Pairwise comparison of the putative 5' regulatory regions of the genes coding for the different isoforms of the Na(+)-K(+)-ATPase catalytic subunit shows the existence of conserved elements, as well as of oligonucleotide blocks with very different structures. It is suggested that the differences in the primary structure of the 5' upstream regions may provide the basis for tissue-specific expression of the Na(+)-K(+)-ATPase isoforms.
Subject(s)
Regulatory Sequences, Nucleic Acid , Sodium-Potassium-Exchanging ATPase/genetics , Amino Acid Sequence , Base Sequence , Binding Sites , Chromosome Mapping , Humans , Isoenzymes , Molecular Sequence Data , Oligonucleotide Probes/genetics , Protein Biosynthesis , Sequence Homology, Nucleic Acid , Transcription Factors/genetics , Transcription, GeneticABSTRACT
The multigene family of human Na,K-ATPase is composed of 5 alpha-subunit genes, 3 of which were shown to encode the functionally active alpha 1, alpha 2 and alpha 3 isoforms of the catalytic subunits. This report describes the isolation, mapping and partial sequencing of the fourth gene (ATP1AL1) that was demonstrated here to be functionally active and expressed in human brain and kidney. Limited DNA sequencing of the ATP1AL1 exons allowed one to suggest that the gene probably encodes a new ion transport ATPase rather than an isoform of the Na,K-ATPase or the closely related H,K-ATPase.
Subject(s)
Adenosine Triphosphatases/genetics , Multigene Family , Sodium-Potassium-Exchanging ATPase/genetics , Transcription, Genetic , Adenosine Triphosphatases/metabolism , Amino Acid Sequence , Base Sequence , Biological Transport , Brain/enzymology , Electrophoresis, Agar Gel , Exons , Humans , Introns , Kidney/enzymology , Molecular Sequence Data , Oligonucleotide Probes , Polymerase Chain Reaction , Sequence Homology, Nucleic AcidABSTRACT
The existence of a chromosome gene family containing at least one gene and one pseudogene was shown for the Na+,K+-ATPase beta-subunit. A partial structure of the beta 1-gene was determined, the coding part of which was completely homologous to cDNA of the Na+,K+-ATPase beta I-subunit from HeLa cells. The region encoding the putative protein transmembrane domain was shown to be bordered by two introns. The structure of a pseudogene (beta psi) was determined. This pseudogene is processed and contains multiple stop codons. Its homology to the beta I-subunit cDNA from HeLa cells is about 88%.
Subject(s)
Sodium-Potassium-Exchanging ATPase/genetics , Animals , Base Sequence , HeLa Cells , Humans , Membrane Proteins/genetics , Molecular Sequence Data , Multigene Family , Pseudogenes , Restriction Mapping , Sequence Homology, Nucleic Acid , SwineABSTRACT
Complete primary structures of both subunits of Na+,K+-ATPase from various sources have been established by a combination of the methods for molecular cloning and protein chemistry. The gene family homologous to the alpha-subunit cDNA of animal Na+,K+-ATPases has been found in the human genome. Some genes of this family encode the known isoforms (alpha I and alpha II) of the Na+,K+-ATPase catalytic subunit. The proteins coded by other genes can be either new isoforms of the Na+,K+-ATPase catalytic subunit or other ion-transporting ATPases. Expression of the genes of this family proceeds in a tissue-specific manner and changes during the postnatal development and neoplastic transformation. The complete exon-intron structure of one of the genes of this family has been established. This gene codes for the form of the catalytic subunit, the existence of which has been unknown. Apparently, all the genes of the discovered family have a similar intron-exon structure. There is certain correlation between the gene structure and the proposed domain arrangement of the alpha-subunit. The results obtained have become the basis for the experiments which prove the existence of the earlier unknown alpha III isoform of the Na+,K+-ATPase catalytic subunit and have made possible the study of its function.
Subject(s)
Biological Evolution , Genes , Sodium-Potassium-Exchanging ATPase/genetics , Amino Acid Sequence , Animals , Base Sequence , Binding Sites , Cell Membrane/enzymology , Humans , Isoenzymes/genetics , Macromolecular Substances , Molecular Sequence Data , Multigene Family , Protein ConformationABSTRACT
The primary structure of the putative regulatory region of a gene of the Na+,K+-ATPase multigene family in the human genome has been determined. This region includes the first exon with all of the untranslatable sequence of mRNA and a dozen nucleotides, coding for the first four amino acids of the hypothetic precursor of the alpha+-subunit. The entire region comprises over 1400 bp. The possible role of specific nucleotide blocks within this region in comparison with other genes is discussed.
Subject(s)
Genes, Regulator , Genes , Multigene Family , Sodium-Potassium-Exchanging ATPase/genetics , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Cosmids , Humans , Mice , Molecular Sequence Data , Rabbits , Restriction Mapping , Sequence Homology, Nucleic AcidABSTRACT
The nucleotide sequence of the rpoB gene of Salmonella typhimurium has been determined in this work. It was compared with known sequences of the gene from other sources and the conservative regions were detected. This allowed some interesting conclusions to be made about the distribution of the functional domains in bacterial RNA polymerase and about the three-dimensional structure of its beta subunit.
Subject(s)
DNA-Directed RNA Polymerases/genetics , Genes, Bacterial , Salmonella typhimurium/genetics , Amino Acid Sequence , Base Sequence , Chloroplasts/analysis , Cloning, Molecular , Escherichia coli/enzymology , Escherichia coli/genetics , Molecular Sequence Data , Plants/genetics , Plants, Toxic , Salmonella typhimurium/enzymology , Sequence Homology, Nucleic Acid , NicotianaABSTRACT
The expression of genes coding for alpha and alpha III isoforms of Na+,K+-ATPase alpha-subunit has been studied in human kidney, brain, thyroid and liver cells. The expression was shown to be subjected to a tissue-specific control and also depended on the developmental stage. The tissue-specific expression of genes coding for different isoforms of the catalytic subunit of Na+,K+-ATPase perhaps may be attributed to various functions of proteins belonging to this family.
Subject(s)
Gene Expression Regulation , Genes , Sodium-Potassium-Exchanging ATPase/genetics , Adult , Aging , Amino Acid Sequence , Base Sequence , Child , Codon , Embryo, Mammalian , Humans , Isoenzymes/genetics , Macromolecular Substances , Molecular Sequence Data , Oligonucleotide Probes , Organ Specificity , RNA, Messenger/geneticsABSTRACT
The BATP gene coding for the beta-subunit of Na+,K+-ATPase has been localized on chromosome 13 of the American mink (Mustela vison) using mink-Chinese hamster somatic cell hybrids and pig cDNA clones as probes. The AATP gene for the alpha-subunit of Na+,K+-ATPase is on mink chromosome 2 [(1987) FEBS Lett. 217, 42-44]. Consequently, the AATP and BATP genes for the Na+,K+-ATPase occupy separate mink chromosomes.
Subject(s)
Mink/genetics , Sodium-Potassium-Exchanging ATPase/genetics , Animals , Cell Line , Chromosome Mapping , Cricetinae , DNA/genetics , Hybrid Cells , Nucleic Acid HybridizationABSTRACT
The primary structure of a gene of the Na+, K+-ATPase multigenic family in the human genome has been determined. The gene corresponds to a hypothetical alpha III-form of the enzyme catalytic subunit. The gene comprises over 25,000 bp, and its protein coding region includes 23 exons and 22 introns. Possible correlation between structural features of the protein and location of introns in the gene are discussed.
Subject(s)
Sodium-Potassium-Exchanging ATPase/genetics , Amino Acid Sequence , Animals , Bacteriophage lambda/genetics , Base Composition , Base Sequence , Binding Sites , Catalysis , DNA/genetics , DNA, Recombinant , Exons , Humans , Introns , Molecular Sequence DataABSTRACT
Intra-individual tissue-specific restriction fragment length polymorphism (RFLP) has been demonstrated in DNA isolated from different mammalian tissues using cDNAs of alpha- and beta-subunits of Na+,K+-ATPase as hybridization probes. We propose that the RFLPs could result from gene rearrangements in the gene loci for the alpha- and beta-subunits of Na+,K+-ATPase. The changes in restriction patterns have been shown to occur during embryonic development and tumor formation. In addition, the tissue specificity of the expression of different genes of the family of Na+,K+-ATPase genes and their low expression in tumor cells have been demonstrated.
