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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 investigated the frequency of rearrangements of the ret and trk proto-oncogenes in Japanese thyroid tumors. DNAs from 38 thyroid papillary carcinomas and 14 follicular adenomas were analyzed by Southern blotting. Rearrangements of the ret and trk proto-oncogenes were detected in one and two papillary carcinomas, respectively, but not in follicular adenomas. Analysis by a
reverse transcriptase
-polymerase chain reaction method showed that the ret rearrangement-positive tumor contained the PTC/retTPC chimeric transcript, which was reported to be found specifically in thyroid tumors and adenomatous
goiter
. We also found that rearranged mRNA of the trk proto-oncogene was expressed at high levels in one of two trk rearrangement-positive tumors. Our results indicated that the frequency of rearrangements of these proto-oncogenes in Japanese papillary carcinomas was much lower than that in Italian patients.
...
PMID:Low frequency of rearrangements of the ret and trk proto-oncogenes in Japanese thyroid papillary carcinomas. 138 40
A
reverse transcriptase
-polymerase chain reaction (RT-PCR) method was adopted for detecting transcripts specific for retTPC/PTC, an activated form of the ret proto-oncogene reported to be found specifically in human papillary thyroid carcinomas. By this sensitive method retTPC/PTC transcript could be detected in about 500 fg of total RNA of TPC-1, a retTPC/PTC transcript-positive cell line. In Japanese patients, one of 11 papillary thyroid carcinomas, four of 19 follicular adenomas and one of two adenomatous goiters were positive for the transcript, indicating that the involvement of retTPC/PTC is not specific to papillary thyroid carcinomas. In several independent RT-PCR experiments using different portions of the same positive carcinoma tissue, retTPC/PTC transcript was always detected. On the other hand, the transcript was not always positive in different RNA samples from benign cases, suggesting that positive carcinomas are probably composed of clonal cell populations all expressing retTPC/PTC, whereas adenomas and adenomatous
goiter
comprise heterogeneous populations: both positive and negative for retTPC/PTC transcript. Activation of the ret proto-oncogene might therefore be involved in malignant conversion to thyroid carcinomas.
...
PMID:Detection of retTPC/PTC transcripts in thyroid adenomas and adenomatous goiter by an RT-PCR method. 171 26
A highly purified thyroglobulin mRNA was isolated from human nodal euthyroid
goiter
. A full-length cDNA was synthesized from 33S RNA by using
reverse transcriptase
in the presence of human placenta ribonuclease inhibitor. A DNA complementary to human Tg mRNA was used in liquid hybridization experiments to quantify Tg mRNA. The amount of Tg mRNA in euthyroid nodal and congenital
goiter
was reduced. In thyroid cancer Tg specific mRNA was absent. Direct correlation between Tg gene expression in thyroid cells and DNAase-I hypersensitivity of chromatin from the thyroid gland nucleus was revealed.
...
PMID:[Changes in the chromatin structure of the thyroid cells related to the expression of the thyroglobulin gene]. 197 92
A highly purified thyroglobulin (Tg) mRNA was isolated from human nodal euthyroid
goiter
. Ultracentrifugation in a 5-20% sucrose density gradient revealed that the protein has a sedimentation coefficient of 33S. cDNA was synthesized from the 33S RNA, using
reverse transcriptase
in the presence of a human placental ribonuclease inhibitor. In order to detect the polymorphism of restriction fragment length, screening of high molecular weight DNA from normal human thyroid gland and from nodal euthyroid and diffuse toxic goiters was carried out, using Tg cDNA probes and plasmid DNA containing a Tg cDNA fragment. After restriction with endonuclease Hind III, three Tg-containing fragments were identified both in the normal and nodal euthyroid goiters, whereas restriction with endonuclease Bam HI revealed two Tg-containing fragments in nodal euthyroid and diffuse toxic goiters.
...
PMID:[Mapping of the thyroglobulin gene in high molecular weight DNA from the human thyroid in normal conditions and in thyroid pathology]. 256 30
Highly purified thyroglobulin mRNA was isolated from human nodal euthyroid
goiter
. Full length cDNA was synthesized from 33S RNA by using
reverse transcriptase
in the presence of human placenta ribonuclease inhibitor. DNA complementary to human Tg mRNA was used in liquid hybridization experiments to determine the quantity of Tg mRNA. The amount of Tg mRNA in euthyroid nodal and congenital
goiter
was reduced. Tg specific mRNA was absent in thyroid cancer cells.
...
PMID:[Thyroglobulin gene expression in human thyroid cells in various types of thyroid pathology]. 258 2
The thyroid gland is a highly vascular tissue, and its blood flow changes dramatically in various pathological conditions. Although the mechanisms regulating these changes in vascularity and blood flow are not well understood, candidate mediators include endothelin-1 (ET-1) and nitric oxide (NO). In the present study, we used a
reverse transcriptase
-polymerase chain reaction assay to determine which components of these vasoregulatory pathways are present in the thyroid and to analyze changes in gene expression in an experimental model of
goiter
formation and involution. Expression of messenger RNAs (mRNAs) encoding ET-1, ET receptors (ETA and ETB), ET-converting enzyme, and the three nitric oxide synthase (NOS) isoforms (NOS I, NOS II, and NOS III) was readily detected in the rat thyroid. After
goiter
formation was induced by thiouracil and a low iodine diet, there was increased expression of the genes encoding ET-related proteins (ET-1, 3.2-fold; ETA, 2.9-fold; ETB, 3.5-fold) as well as two of the three NOS isoforms (NOS I, 2.7-fold; NOS III, 4.9-fold). During iodide-induced involution, the ET-related mRNA levels remained elevated, whereas those of the two NOS isoforms returned to basal values. ET-converting enzyme, NOS II, and thyroglobulin mRNAs were minimally affected in this model, providing evidence for selective regulation of these genes. To assess whether NO plays a role in vascular changes during
goiter
formation, animals were treated with a NOS inhibitor, N-nitro-L-arginine methyl ester (NAME). NOS activity in the thyroid was inhibited by more than 75% after treatment with NAME. Thyroid hormone and TSH levels were unchanged. Although NAME had little effect on overall thyroid size, vascular expansion during
goiter
formation was decreased by 36%. We conclude that the thyroid gland expresses a complex network of vasoactive genes whose expression is regulated dynamically during thyroid
goiter
formation and involution. NO production and probably other locally produced vasoactive substances are involved in changes in thyroid vascularization.
...
PMID:Expression of nitric oxide synthase isoforms in the thyroid gland: evidence for a role of nitric oxide in vascular control during goiter formation. 758 72
TTF-1 and Pax-8 are thyroid-specific transcription factors, from homeo and paired box genes, respectively, that are responsible for thyroid development and for thyroglobulin and thyroperoxidase gene expression. However, TTF-1 and Pax-8 preferentially bind to the thyroglobulin and thyroperoxidase promoters, respectively. Here, we have studied a patient with defective thyroglobulin synthesis. Thyroglobulin mRNA was found at very low levels while the mRNA for thyroperoxidase was found to be more abundant compared with control tissue. The low levels of thyroglobulin mRNA are caused by a transcriptional defect due to the virtual absence of TTF-1 expression as determined by Northern blot analysis,
reverse transcriptase
-PCR, and electrophoretic mobility shift assays. The level of Pax-8 mRNA was the same in the
goiter
and in the control thyroid. These results are the first reported evidence of a congenital
goiter
with a thyroglobulin synthesis defect due to the low expression of the thyroid-specific transcription factor TTF-1. Moreover, these data suggest that TTF-1 and Pax-8 would be differentially regulating thyroglobulin and thyroperoxidase gene transcription.
...
PMID:Congenital human thyroglobulin defect due to low expression of the thyroid-specific transcription factor TTF-1. 763 72
Restricted expression of oncofetal fibronectin mRNA in the tissues of thyroid papillary and anaplastic carcinoma has recently been shown by both Northern blot analysis and
reverse transcriptase
polymerase chain reaction (RT-PCR). Oncofetal fibronectin mRNA can be a target of gene diagnosis and targeted gene therapy, provided it is expressed in all cancer cells in the tissues. To investigate this criterion in thyroid cancer tissues, we measured their expression of oncofetal fibronectin mRNA using in situ hybridization. An abundant expression of oncofetal fibronectin mRNA was found in all the observed cancer cells of six papillary carcinomas and an anaplastic carcinoma, but not in the tissues of normal thyroid, Graves' disease, adenomatous
goitre
, follicular adenoma, follicular carcinoma or medullary carcinoma. This result encourages us to establish gene diagnosis of thyroid papillary and anaplastic carcinomas by detecting oncofetal fibronectin mRNA in biopsies.
...
PMID:Restricted expression of oncofetal fibronectin mRNA in thyroid papillary and anaplastic carcinoma: an in situ hybridization study. 968 97
The ability to concentrate iodide, a fundamental property of normally functioning thyroid tissue, is altered in various thyroid diseases. Given the critical role of the Na+/I- symporter (NIS) in controlling iodide access to the thyroid gland, altered expression of NIS may be responsible, at least in part, for an enhanced or diminished capacity to concentrate iodide. In this study, we used Northern blot analysis, a newly established quantitative polymerase chain reaction (PCR) assay and in addition hNIS-directed immunohistochemical analysis to assess the levels of hNIS mRNA and protein expression in various localized and diffuse benign thyroid abnormalities, including Graves' disease (GD), scintigraphically cold solitary benign thyroid nodule (CBTN), nontoxic multinodular goiter (NMNG), solitary autonomously functioning thyroid nodule (AFTN), and mild diffuse iodine deficiency
goiter
(IDG). In addition, in view of the recent identification of putative binding sites for the transcription factors thyroid transcription factor-1 (TTF-1) and human paired-box-protein-8 (Pax-8) in the human NIS gene promoter, we used
reverse transcriptase
-polymerase chain reaction (RT-PCR) to assess in these same samples the levels of TTF-1 and Pax-8 gene expression. Northern blot analysis revealed high levels of hNIS gene expression in thyroid specimens derived from patients with GD and AFTN. In contrast, levels of hNIS mRNA expression were moderate in NMNG, low in diffuse IDG, and very low in CBTN. Quantitative RT-PCR analysis of hNIS mRNA transcripts revealed variable but generally low levels of hNIS gene expression in IDG and NMNG, and undetectable or very low levels of hNIS mRNA in all scintigraphically CBTN studied. In contrast, markedly elevated levels of hNIS mRNA transcripts were detected in active GD (up to 17-fold) and AFTN (up to 25-fold). Immunohistochemical analysis revealed abundant hNIS protein expression by thyroid follicular cells in GD, moderate and heterogeneous levels in NMNG, and very low levels in CBTN. hNIS mRNA levels were correlated with TTF-1 and Pax-8 gene expression in GD and, to a lesser degree, in AFTN, NMNG, and IDG, but not in CBTN. In general, hNIS gene expression was more closely correlated with TTF-1 as compared to Pax-8 gene expression. In conclusion, the abundance of hNIS mRNA and protein expression in a broad range of benign thyroid pathologies correlated well with their functional state as assessed by thyroid scintigraphy. In addition to TTF-1 and Pax-8, other transcription factors and enhancer elements may contribute to regulation of NIS gene promoter activity.
...
PMID:Analysis of human sodium/iodide symporter, thyroid transcription factor-1, and paired-box-protein-8 gene expression in benign thyroid diseases. 1036 77
Resistance to thyroid hormone (RTH) is a clinical syndrome characterized by elevated serum thyroid hormone (TH) levels, unsuppressed thyrotropin (TSH) levels, and tissue hyposensitivity to TH. In almost all cases, the genetic basis of RTH lies in mutation of one of the two TH receptor beta (TRbeta) alleles. Recently, patients from several families with phenotypic manifestations of RTH in the absence of TR mutations have been described. We report a case of a 31-year-old woman who presented with
goiter
, tachycardia, elevated TH levels, unsuppressed TSH, and "inappropriately normal" levels of peripheral TH action markers. In two separate clinical evaluations, the patient exhibited typical clinical and biochemical evidence for peripheral and pituitary RTH. Surprisingly,
reverse transcriptase
-polymerase chain reaction (RT-PCR) of full-length TRalpha and TRbeta mRNAs, and genomic PCR using primers flanking exons encoding the carboxy-terminal region of TRbeta failed to demonstrate mutations in the TRalpha or TRbeta genes. It is likely that defects in the regulation of TR genes or mutations in transcriptional cofactors involved in TR signaling account for this patient's phenotype.
...
PMID:Resistance to thyroid hormone in a patient without thyroid hormone receptor mutations. 1183 36
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