Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have previously reported functional alterations in vitro in the hematopoietic compartment of patients with diffuse large B-cell lymphoma (DLBCL). In the present study, we assessed the presence of molecular alterations in hematopoietic cells derived from DLBCL marrow. Accordingly, the expression of four genes (i.e. ice, bcl-2, c-myc and p53) was assessed both, at the mRNA and protein levels, in three cell populations: (i) population I, consisting of morphologically recognizable precursor and mature cells; (ii) population II, enriched for CD34+ Lineage-negative (Lin-) cells; and (iii) population III, enriched for CD34+ CD38- Lin- cells. By using a multiplex reverse transcriptase-polymerase chain reaction system, we observed reduced expression of bcl-2 in population I, and c-myc in populations I and II from lymphoma marrow compared to their normal counterparts. On the other hand, expression of ice and p53 was not significantly different when comparing normal and DLBCL samples. At the protein level, all four molecules were expressed in a higher proportion of samples from DLBCL patients than in marrow samples from normal subjects. Interestingly, these proteins were expressed predominantly in primitive cells (population III), whereas the proportion of positive samples was reduced in population II, and even more in population I. Taken together, our results indicate that, in DLBCL, molecular alterations are present in hematopoietic cells from bone marrow, including morphologically recognizable precursor and mature cells, as well as primitive hematopoietic progenitors (CD34+ cells). To date, the physiological implications of these alterations are still unclear, and further studies should be undertaken to address this issue.
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PMID:Expression of ice, bcl-2, c-myc and p53 in different bone marrow cell populations from patients with diffuse large B-cell lymphoma. 1669 May 25

Colorectal carcinogenesis is initiated mainly by aberrant activation of the Wnt signaling pathway, caused by mutation of either APC or beta-catenin (CTNNB1) gene. Poly(ADP-ribose) polymerase-1 (PARP-1) is a highly conserved nuclear enzyme, which binds tightly to DNA and plays a role in DNA repair, recombination, proliferation and genomic stability. It has recently been shown that PARP-1 is a novel co-activator of TCF-4/beta-catenin-evoked gene transactivation and may play a role in colorectal carcinogenesis. The aim of this study was to examine the PARP-1 expression and determine whether it is correlated with the expression of beta-catenin and its target genes such as c-myc, cyclin D1 and matrix metalloproteinase (MMP)-7 in the early stage of sporadic colorectal carcinogenesis. Using the semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), 91 colorectal tumours, including 65 adenomas and 26 submucosal (pT1) cancers, were analysed for the expression of PARP-1, beta-catenin, c-myc, cyclin D1 and MMP-7. Immunohistochemical analysis of PARP-1 and beta-catenin was also performed. PARP-1 mRNA overexpression was detected in 64 (70.3%) of the 91 tumours. PARP-1 overexpression was significantly correlated with tumour size and histopathology. Overexpression of beta-catenin, c-myc, cyclin D1 and MMP-7 mRNA expression was observed in 39.6%, 78.0%, 83.5% and 72.5% of the 91 tumours, respectively. PARP-1 overexpression was correlated significantly with overexpression of beta-catenin, c-myc, cyclin D1 and MMP-7. Correlation of PARP-1 expression with beta-catenin overexpression was also demonstrated by immunohistochemistry. The results suggest that PARP-1, in conjunction with beta-catenin, c-myc, cyclin D1 and MMP-7, plays an important role in the early stage of colorectal carcinogenesis.
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PMID:Overexpression of poly(ADP-ribose) polymerase-1 (PARP-1) in the early stage of colorectal carcinogenesis. 1680 31

Caffeic acid phenethyl ester, an active component of propolis, has been implicated in the regulation of cell growth and apoptosis, although the exact mechanism of this activity has not been elucidated. In this study, we explored the effects of caffeic acid phenethyl ester on growth, cell cycle, apoptosis and beta-catenin/T-cell factor signaling in human colon cancer cells. Using two human sporadic colon cancer cell lines (HCT116 and SW480), we assayed for cell growth inhibition, cell cycle and apoptosis induction. We also assayed for beta-catenin and downstream target genes (cyclin D1 and c-myc) mRNA and protein expression by reverse transcriptase-polymerase chain reaction and Western blot analysis. Beta-catenin localization was detected by indirect immunofluorescence. Beta-catenin/T-cell factor transcriptional activity was determined by transient transfection and reporter gene assay. Caffeic acid phenethyl ester completely inhibited growth, and induced G1 phase arrest and apoptosis in a dose-dependent manner in both HCT116 and SW480 cells. Treatment of human colon cancer cells with apoptotic concentrations of caffeic acid phenethyl ester resulted in a dose-dependent and time-dependent loss of total beta-Catenin protein, associated with decreased nuclear beta-catenin. Caffeic acid phenethyl ester reduced the expression of cyclin D1 and c-myc in a dose-dependent and time-dependent manner. We proved that caffeic acid phenethyl ester markedly suppressed the transcriptional activity of beta-catenin/T-cell factor in both HCT116 and SW480 cells depending on the concentration of caffeic acid phenethyl ester. These results indicate that caffeic acid phenethyl ester is an excellent inhibitor of beta-catenin/T-cell factor signaling in colon cancer cell lines and suggest that caffeic acid phenethyl ester merits further study as an agent against colorectal cancers.
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PMID:Caffeic acid phenethyl ester induces growth arrest and apoptosis of colon cancer cells via the beta-catenin/T-cell factor signaling. 1692 25

Molt-inhibiting hormone (MIH), a member of the crustacean hyperglycemic neuropeptide hormone family, inhibits ecdysteroidogenesis in the molting gland or Y-organ (YO). A cDNA encoding MIH of the land crab (Gel-MIH) was cloned from eyestalk ganglia (EG) by a combination of reverse transcriptase polymerase chain reaction (RT-PCR) and 3'- and 5'-rapid amplification of cDNA ends (RACE). The cDNA (1.4 kb) encoded MIH prohormone containing a 35 amino acid signal peptide and a 78 amino acid mature peptide. The mature peptide had the six cysteines, one glycine, two arginines, one aspartate, one phenylalanine, and one asparagine in identical positions in the highly conserved sequence characteristic of other crustacean MIHs. Gel-MIH was expressed only in the EG, as determined by RT-PCR; it was not detected in Y-organ, heart, integument, gill, testis, ovary, hepatopancreas, thoracic ganglion, or skeletal muscle. A cDNA encoding the mature peptide was used to express recombinant MIH (rMIH) using a yeast (Pichia pastoris) expression system. Two constructs were designed to yield either a mature MIH fusion protein with a c-myc epitope and histidine (His) tag at the carboxyl terminus or an untagged mature protein without the c-myc and His sequences. Immunoreactive peptides were detected in Western blots of the cell culture media with both MIH constructs, indicating secretion of the processed rMIH into the medium. Culture media containing the untagged mature peptide significantly inhibited ecdysteroid secretion by YOs from land crab and green crab (Carcinus maenas) cultured in vitro, indicating that the Gel-rMIH was biologically active.
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PMID:Molt-inhibiting hormone from the tropical land crab, Gecarcinus lateralis: cloning, tissue expression, and expression of biologically active recombinant peptide in yeast. 1709 91

Antisense therapy is limited in application in clinical therapy because of two existing problems: rapid degradation of antisense nucleic acids and poor diffusion across the cell membrane. Here we report the use of single-walled carbon nanotubes as delivery system for transporting antisense myc into HL-60 cells. Antisense-myc-conjugated single-walled carbon nanotubes were synthesized, characterized by atomic force microscopy (AFM), fluorescent microscopy, and Raman spectroscopy. Incubated with HL-60 cells at 37 degrees C, the single-walled carbon nanotubes (SWNTs) inside HL-60 cells were measured quantitatively by HPLC. Expression of c-myc gene and protein were analyzed by reverse transcriptase PCR and Western blot, respectively. Results showed that as-myc-conjugated SWNTs dropped character peaks at quadruple wavenumber (cm(-1)) compared with SWNTs, and could enter into HL-60 cells within 15 min after incubation, whose uptake amounts by HL-60 cells increased as the incubation time increased. After 48 hours, the amount of SWNTs in HL-60 cells began to decrease. Compared with as-myc and SWNTs, as-myc-conjuagted SWNTs exhibited the strongest inhibition on the proliferation of HL-60 cells, induced cell apoptosis, and down-regulated lowest expression of c-myc gene and C-MYC protein. The SWNTs can directly deliver as-myc into HL-60 cells, enhance the inhibition of as-myc on HL-60 cells, and is likely a better delivery system for antisense therapy. Antisense modified SWNTs can be used for intracellular gene regulation with potential applications in tumor therapy and drug delivery.
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PMID:Effects of antisense-myc-conjugated single-walled carbon nanotubes on HL-60 cells. 1745 Sep 37

The presence of a complex population of mRNAs in human mature spermatozoa is well documented; among them, transcripts of aromatase and ERs (oestrogen receptors) have been described but their significance is not clear. Therefore, to clarify the role of this complex population of mRNAs in human ejaculated sperm, we have isolated on discontinuous density gradients two main fractions from the same sample: high- and low-motile spermatozoa. The levels of different transcripts coding for molecules involved in nuclear condensation [Prm-1 (protamine 1) and Prm-2], capacitation [eNOS (endothelial nitric oxide synthase), nNOS (neuronal nitric oxide synthase), c-myc], motility and sperm survival (aromatase) have been assessed using semi-quantitative RT (reverse transcriptase)-PCR. The viability of sperm as well as the percentage of apoptosis were identical in high- and low-motile fractions. No significant change in the c-myc/Prm-2 ratio between the two populations of spermatozoa was observed. Conversely the amount of Prm-1 mRNA was significantly higher in low-motile than in high-motile fraction; in most of the high-motile sperm samples analysed, eNOS and nNOS transcripts were undetectable, whereas they were observed in low-motile sperm. Moreover, a partial or complete disappearance of c-myc transcripts was observed after capacitation. As to the aromatase expression, a significant decrease in the amount of transcripts in immotile sperm fraction was recorded in all samples studied. To conclude, analysing mRNA profiles in humans could be helpful either as a diagnostic tool to evaluate male fertility, since they reflect spermatogenesis gene expression, and/or a prognosis value for fertilization, since these RNAs are delivered to oocytes.
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PMID:RNA dynamics of fertile and infertile spermatozoa. 1751 68

Pineal parenchymal tumours (PPT) are rare neoplasms and there have been few in vitro studies. Their capacity for synthesizing and secreting melatonin has been only partially examined. We investigated the presence of messenger RNA (mRNA) encoding tryptophan hydroxylase (TPH), arylalkylamine N-acetyltransferase (AANAT), hydroxyindol-O-methyltransferase (HIOMT), three enzymes involved in melatonin synthesis, and c-myc, a tumoural marker, in 10 PPT, one papillary tumour of the pineal region (PTPR), cell cultures derived from four PPTs and from three other tumours of the pineal region, and in normal pineal gland. Moreover, protein expression of TPH was investigated in three PPT and PTPR. Quantitative real-time reverse transcriptase-polymerase chain reaction and immunohistochemistry were used and the melatonin production by tumoural cells in vitro was analysed by radioimmunoassay. We showed that all the tumoural tissues and cells contained c-myc mRNA. mRNAs encoding TPH, AANAT and HIOMT were detected in all PPT, suggesting that tumour cells can synthesize melatonin. Only PPT expressed TPH protein. Cultured cells lost expression of transcripts throughout passages even if ultrastructural study revealed the presence of characteristic organelles in these tumoural cells. Nevertheless, the basal secretion of melatonin observed in one PPT culture is in favour of a maintained melatonin production and secretion by tumoural pinealocytes, but melatonin production was not stimulated by a beta noradrenergic agonist. Moreover, PTPR never expressed mRNA encoding TPH, AANAT and HIOMT. Our results may contribute to a better understanding of the biology of PTT and PTPR and may help to the diagnosis of these rare tumours.
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PMID:Histological features and expression of enzymes implicated in melatonin synthesis in pineal parenchymal tumours and in cultured tumoural pineal cells. 1797 Oct 73

An assay for rapid and direct detection of DNA/mRNA using cationic fluorescent polymer based on one-dimensional microfluidic beads array (1-D chip) has been developed. The cationic water-soluble polythiophene derivatives can easily transduce hybridization events into measurable optical signal due to the conformational changes of the conjugated backbone, when mixed with single-stranded or double-stranded oligonucleotides. In this paper, the polymer was introduced into 1-D chip for fluorescence detection of nucleic acids, and demonstrated its capability on rapid detection of p53 complementary DNA (cDNA) with different concentration. Using this system, we have evaluated the mRNA expression changes of three tumor-associated genes (p53, c-myc and cyclin-d1) in human nasopharyngeal carcinoma CNE2 cell lines before and after 5-flouorouracil (5-FU) stimuli. These results were validated by the conventional reverse transcriptase-PCR. The most important advantage of this assay is not needed target or report labeling prior to hybridization, which makes the experiment process easy to handle and low-cost for multi-target measurement.
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PMID:Nucleic acids detection using cationic fluorescent polymer based on one-dimensional microfluidic beads array. 1906 86

Mesenchymal stromal cells (MSCs), seeded onto a scaffold and associated with platelet-gel, may represent an innovative treatment to improve bone repair. The preparation of MSCs for clinical use requires the fulfillment of Good Manufacturing Practice indications. The aim of this study was to validate a Good Manufacturing Practice-grade protocol of tissue engineering for bone regeneration, seeding platelet lysate (PL)-cultured MSCs onto an hydroxyapatite clinical-grade scaffold. Six large-scale experiments were performed. MSC expansions were performed comparing fetal bovine serum 10% and PL 5%. We demonstrated that PL lots contain high levels of growth factors possibly responsible of accelerated growth rate, since the number of colony-forming unit-fibroblast and population doublings were always significantly higher in PL cultures. MSCs were characterized for their phenotype and multilineage differentiation capacity, demonstrating appropriate features for both conditions. Gene expression analysis revealed higher expression of typical osteogenic genes of PL-cultured MSCs, when compared to fetal bovine serum MSCs. Cell transformation was excluded by analysis of karyotype, absence of growth without anchorage, and p53/c-myc gene expression. Scaffolds were precoated with retronectin before MSC seeding. MSC adhesion, distribution, and proliferation were demonstrated through the whole surface of the scaffold by scanning electron microscopy analysis or by immunofluorescence and MSC osteogenic differentiation through quantitative reverse transcriptase-polymerase chain reaction of typical osteogenic genes. The present report offers a model of an MSC-based bioengineered device, using an hydroxyapatite clinical-grade scaffold, and supports its potential use in tissue engineering to repair bone defects.
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PMID:Characterization of platelet lysate cultured mesenchymal stromal cells and their potential use in tissue-engineered osteogenic devices for the treatment of bone defects. 1946 94

The study was purposed to explore the expressions of pituitary tumor transforming gene and c-myc gene in patients with multiple myeloma (MM) and its relationship with pathogenesis of MM. Expressions of pituitary tumor transforming gene and c-myc gene mRNA in BMMNC from 33 patients with MM and 10 normal controls were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that the expressions of pituitary tumor transforming gene and c-myc gene mRNA were significantly higher in MM patients those that in normal controls (p<0.05). The expression of pituitary tumor transforming gene mRNA was significantly correlated with the expression of c-myc gene (r=0.801, p<0.05). In conclusion, the overexpressions of pituitary tumor transforming gene and c-myc gene may be related to the pathogenesis and progression of MM.
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PMID:[Expression of pituitary tumor transforming gene and C-myc gene in patients with multiple myeloma]. 1984 Apr 58


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