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)

Interleukin (IL)-7 transgenic mice, which we established previously, developed severe dermatitis characterized by massive infiltration of gammadelta T cells in the dermal lesion. To fully understand the pathology of this intriguing skin disease, we examined several immunologic features of dermis infiltrating lymphocytes from the lesional skin of IL-7 transgenic mice. We observed a moderate response to mitogens, a poor response to alloantigens, and the absence of cytotoxic activities to several tumor cell lines and skin derived cells regardless of the presence of IL-2 or IL-7. On the other hand, dermis infiltrating lymphocytes could proliferate with exogenous IL-2 and IL-7. Moreover, reverse transcriptase polymerase chain reaction and fluorescence activated cell sorter analysis revealed that dermis infiltrating lymphocytes expressed various cytokines including IL-4 and IL-7, and several activation markers for T cells (CD44, CD69, IL-2R alpha), in addition to IL-7R alpha. In the sera of the affected mice, hyper epsilon-globulinemia was observed. These findings suggested that dermis infiltrating lymphocytes proliferated in an activated state in the skin lesion in an autocrine and/or paracrine manner and produced Th2 type cytokines that might evoke immunologic abnormalities. This study and previous findings suggest that IL-7 transgenic mouse with dermatitis offer the potential of serving as a useful tool for investigating the immunologic role of cutaneous gammadelta T cells, especially their participation in IgE production in vivo.
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PMID:Immunologic abnormalities exhibited in IL-7 transgenic mice with dermatitis. 957 38

The aim of this study was to investigate certain genes for their suitability as molecular markers for detection of breast carcinoma cells using the reverse transcriptase-polymerase chain reaction (RT-PCR). RNA was prepared from MCF-7 breast carcinoma cells and peripheral blood leucocytes of healthy female volunteers. This RNA was screened for mRNA of MUC1, cytokeratin 19 (CK19) and CD44 (exons 8-11) by RT-PCR and the results validated by Southern blots. Variable degrees of expression of MUC1 and CD44 (exons 8-11) were detected in normal peripheral blood, rendering these genes non-specific for epithelial cells and therefore unsuitable for use as markers to detect breast carcinoma cells. Although CK19 mRNA was apparently specific, it was deemed unsuitable for use as a marker of breast cancer cells in light of its limited sensitivity. Furthermore, an attempt at using nested primers to increase sensitivity resulted in CK19 mRNA being detected after two amplification rounds in blood from healthy volunteers.
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PMID:Putative markers for the detection of breast carcinoma cells in blood. 957 23

Despite recent extensive immunohistochemical studies, the expression patterns of CD44 in testicular germ cell tumors are still controversial. In the present study, we investigated the CD44 gene expression in 40 specimens including 18 seminomas, 16 nonseminomatous germ cell tumors (NSGCT), and 6 normal testes by reverse transcriptase-polymerase chain reaction and Western blotting. Reverse transcriptase-polymerase chain reaction analysis revealed that the standard CD44 isoform (CD44s) was expressed in all of the specimens, whereas the variant CD44 isoforms were highly expressed in NSGCTs but barely detectable in seminomas and normal testes. In addition, we confirmed by direct DNA sequence analysis that the predominantly expressed variant isoform in NSGCTs was CD44v8-10. In germ cell tumors, these results were paralleled in Western blot analysis; that is, CD44s protein was expressed in all of the tumor specimens, whereas high molecular weight variant isoforms were expressed only in NSGCTs. However, at the protein level, no detectable CD44 was expressed in normal testes. These findings show that the combined assessment of CD44 expression patterns at both the RNA and protein levels enables us to distinguish among seminoma, NSGCT, and normal testis specimens; hence, it could serve as a useful practical adjunct to conventional diagnostic methods for testicular germ cell tumors.
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PMID:Expression patterns of CD44 adhesion molecule in testicular germ cell tumors and normal testes. 958 83

In the present study, we have employed a unique breast cancer cell line (Met-1, which was derived from a high metastatic potential tumor in transgenic mice expressing polyomavirus middle T oncogene) to study the role of CD44 variant isoform(s) in the regulation of metastatic breast tumor cell behavior. The results of reverse transcriptase-polymerase chain reaction, Southern blot, nucleotide sequencing, immunoprecipitation, and immunoblot analyses indicated that these cells express a major CD44 isoform (molecular weight approximately 260 kDa) containing a v3,8-10 exon insertion (designated as CD44v3,8-10). In addition, we have determined that CD44v3,8-10 binds specifically to the cytoskeletal proteins such as ankyrin. Biochemical analyses, using competition binding assays and a synthetic peptide identical to NGGNGTVEDRKPSEL (a sequence located between aa480 and aa494 of CD44v3,8-10) indicate that this 15-amino acid peptide binds specifically to the cytoskeletal protein ankyrin (but not to fodrin or spectrin). This peptide competes effectively for ankyrin binding to CD44v3,8-10. Therefore, we believe that the sequence 480NGGNGTVEDRKPSE494L, located at the cytoplasmic domain of CD44v3,8-10, is required for the ankyrin binding. We have also detected that CD44v3,8-10-containing Met-1 cells are capable of forming membrane spikes or "invadopodia" structures and undergo active migration processes. Treatments of Met-1 cells with certain agents including anti-CD44v3 antibody, cytochalasin D (a microfilament inhibitor), and W-7 (a calmodulin antagonist), but not colchicine (a microtubule disrupting agent) effectively inhibit "invadopodia" formation and subsequent tumor cell migration. Further analyses using zymography assays and double immunofluorescence staining indicated that CD44v3,8-10 is closely associated with the active form of matrix metalloproteinase, MMP-9, in a complex within "invadopodia" structures. These findings suggest that CD44v3,8-10 plays an important role in linking ankyrin to the membrane-associated actomyosin contractile system required for "invadopodia" formation (coupled with matrix degradation activities) and tumor cell migration during breast cancer progression.
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PMID:CD44v(3,8-10) is involved in cytoskeleton-mediated tumor cell migration and matrix metalloproteinase (MMP-9) association in metastatic breast cancer cells. 961 60

We examined the expression of selected growth factors, growth factor receptors, elements of extracellular matrix and cell adhesion molecules in the germinal matrix layer (GML) utilizing immunohistochemistry and reverse transcriptase polymerase chain reaction. At autopsy brain samples from 10 neonatal infants were used. Epidermal growth factor receptor (EGFR) was significantly expressed in the matrix cells. While transforming growth factor alpha and heparin-binding epidermal growth factor-like growth factor were found in the matrix cells or vascular wall as ligands, epidermal growth factor was not expressed. EGFR and its ligands are thought to be important factors for the maintenance of the matrix cells and cell-to-cell interactions. Insulin like growth factor I, its receptor Ibeta and tenascin were found in the stroma of the GML and periventricular region. Vascular endothelial growth factor and receptor Flk-1, laminin A and B2, fibronectin, collagen type IV and integrins such as beta3, alpha5beta1 and alphaVbeta3 were found mainly in or around the vascular wall indicating their important roles for vascularization. Transforming growth factor beta2 and its receptor II were expressed in the matrix cells and/or vascular wall suggesting a role in proliferation and/or regression of the vasculature. CD44 and Thy-1 were also expressed in the matrix cells.
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PMID:Growth factors in infant germinal matrix: relationship to extracellular matrix and cell adhesion molecules. 973 49

CD44 forms a group of transmembranous glycoproteins formed by alternative splicing of a single mRNA. The expression of v6 exon-containing variants correlates with metastasis and poor prognosis in a number of malignancies. The distribution and prognostic value of CD44s, CD44v5, and CD44v6 were studied immunohistochemically in the radical prostatectomy specimens of 97 patients with prostate cancer and in 12 lymph node metastases. The mean follow-up period was 84 months. The percentage of CD44-immunoreactive cells was scored semiquantitatively. CD44 mRNA expression was studied in nine prostate cancer and eight benign prostatic hyperplasia (BPH) samples by reverse transcriptase-PCR. Benign prostatic glands almost always expressed CD44s, CD44v6, and, at a lower intensity, CD44v5. CD44 scores decreased from low- to high-grade prostatic intraepithelial neoplasia. CD44s, CD44v5, and CD44v6 were expressed in 86, 23, and 69% of the adenocarcinomas, respectively. Gleason sum score (GSS) and pT stage were correlated inversely with CD44s and CD44v6 scores. CD44 was not found in the lymph node metastatic tumor cells. At the mRNA level, 89% of the tumors and all BPH samples expressed CD44s. CD44v6-v10 mRNA was present in 44 and 75% of the tumors and BPH samples, respectively. Loss of CD44s and CD44v6 predicted an adverse prognosis at univariate analysis. The independent prognosticators identified by multivariate analysis were: GSS, pT stage, and CD44s for clinical progression; GSS and CD44s for prostate-specific antigen progression; and GSS for tumor-specific survival. Loss of CD44s expression in prostate adenocarcinoma predicts a poor prognosis, independent of stage and grade.
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PMID:The prognostic value of CD44 isoforms in prostate cancer patients treated by radical prostatectomy. 981 53

In a previous report, we investigated inflammatory responses induced by injecting Listeria monocytogenes into one testis of a mouse. We demonstrated that the contralateral testis also developed an orchitis despite the absence of bacteria, indicating that the inflammation on the uninfected, contralateral side was of autoimmune character. In both infected and autoimmune testes, gammadelta and alphabeta T cells infiltrated during the inflammation. In this paper, we present the data of a comparison of the character of gammadelta T cells of the infected and autoimmune testes. In both testes, gammadelta T cells appeared to be activated, as assessed by high CD44 and low l-selectin expression. Analysis of T-cell receptor (TCR) usage in both inflammation types revealed the same gammadelta TCR repertoire. Finally, the semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrated that gammadelta T cells in both types of inflammation were capable of producing interleukin-2 (IL-2), IL-4, interferon-gamma (IFN-gamma), IL-10 and transforming growth factor-beta (TGF-beta). These results imply that gammadelta T cells present in infected-induced and autoimmunity-induced inflammation have the same characteristics and could work as immunoregulatory cells.
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PMID:Gamma delta T cells in infection-induced and autoimmune-induced testicular inflammation. 982 3

CD44 isoforms belong to a family of cell adhesion molecules expressed on the cell surface of many tumor cells during human breast cancer progression. In this study we have analyzed the expression of CD44v3-containing isoforms [containing heparan sulfate addition sites for growth factor binding] in primary breast tumors, axillary nodal metastases and normal breast tissue. Using reverse transcriptase-polymerase chain reaction (RT-PCR) followed by Southern blot, cloning, nucleotide sequencing and RT-in situ-PCR analyses, we have found that at least two CD44v3-containing isoforms, including one new species of CD44v2,deltav3-10 (deltav3 defined as a v3 exon lacking the first 24 base pairs) and another previously reported CD44v3,8-10 are preferentially expressed in human primary breast tumor and axillary nodal metastases but not in normal breast tissues. These finding suggest that these CD44v3-containing isoforms are closely associated with breast cancer metastasis.
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PMID:A new CD44V3-containing isoform is involved in tumor cell growth and migration during human breast carcinoma progression. 987 31

CD44v6 expression appears to be associated with adverse prognosis and propensity for metastasis in patients with colorectal cancer. However, expression of CD44 variants in different tumour stages has been poorly characterised. CD44 variant expression was investigated in normal colonic mucosa (n = 36), colorectal adenomas (n = 15), carcinomas (n = 62) and metastases (n = 6) by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blotting with exon-specific probes. High frequencies of CD44 standard (CD44s) and CD44 epithelial (CD44e) were observed in normal and neoplastic tissue. CD44v2 was seen predominantly in adenomas (27%) and UICCI carcinomas (29%). CD44v5 expression was low in normal mucosa (3%), higher in adenomas and carcinomas (29-33%), independent of tumour stage. CD44v6 expression was low in normal mucosa (6%) and higher in adenomas (47%) and carcinomas (42%). Surprisingly, a significant decrease of CD44v6 was observed in metastatic primary tumours (8%) and metastases (17%) (UICCIV) (P < or = 0.05). Therefore, the concept of CD44v6 conferring metastatic potential to malignant cells cannot be supported by our data.
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PMID:Decreased expression of CD44 splicing variants in advanced colorectal carcinomas. 989 37

The purpose of this study was to investigate whether CD44v6 expression correlates with progression or metastasis of cervical cancer. The presence of mRNA for CD44v6 was examined, the association with clinicopathological features was assessed in 80 patients with cervical cancer by reverse transcriptase-polymerase chain reaction (RT-PCR) and subsequent Southern blot hybridisation with an oligonucleotide probe specific for v6. The standard form of CD44 was expressed in all specimens and 53 of 80 cervical cancers expressed an isoform containing exon v6 in combination with other variant exons. In addition, longer size transcripts of more than 1350 bp (long form) were identified in 22 of the 53 CD44v6 positive patients. The expression of CD44v6 and CD44v6 long form in squamous cell carcinomas was significantly higher than that in non-squamous cell carcinomas (P < 0.001). The expression of CD44v6 long form in histological grade 1 and 2 was significantly higher than that in grade 3 (P < 0.05). 47 patients in stage Ib-IIb cervical cancers were treated by radical hysterectomy and pelvic lymphadenectomy. We did not find any association between the expression of the long form or the short form of CD44v6 and any pathological features, except for histological cell type. These findings suggest that the regulation of CD44v6 seems to be different between different histological cell types and different tumour grades, and the expression of CD44v6 might not be implicated in the progression and metastasis of cervical cancer.
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PMID:CD44 exon v6 correlates with cellular differentiation but not with progression and metastasis of cervical cancer. 1007 Mar 19


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