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 examined the expression of alpha 1, alpha 2, alpha 3, alpha 4, alpha 5 and beta 1 integrin on 36 transitional cell cancers (TCCs) in the bladder by immunohistochemistry. Only alpha 2, alpha 3 and beta 1 were detected on normal transitional cell epithelium, but four TCCs (12.5%) revealed positive staining for alpha 1, seven (19.4%) for alpha 4 and seven (20%) for alpha 5. These altered expressions of integrin alpha chain were more frequent in histologically higher stage or grade of TCC, and a correlation was found between increased alpha 5 expression and histological stage. alpha 5 was positive in 6 (35.3%) of 17 invasive TCCs whereas only 1 (5.9%) of 17 superficial TCCs. Flow cytometric analysis on bladder cancer cell lines showed that T24 and HT1376, which are undifferentiated TCC cell lines, highly expressed alpha 5 and beta 1. Also, SCaBER, which is derived from urinary bladder squamous cell cancer and which is recognised as the most malignant phenotype after metaplasia of transitional epithelium, had alpha 5 and beta 1. However, RT4, which is derived from transitional cell papilloma, showed no expression of alpha 5. Furthermore, reverse transcriptase-polymerase chain reaction (RT-PCR) showed the presence of mRNA of alpha 5 on T24, SCaBER and HT1376, but not on RT4. Taken together, it seems that the presence of alpha 5 integrin might be a more malignant phenotype in transitional cell carcinoma.
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PMID:Correlation between integrin alpha 5 expression and the malignant phenotype of transitional cell carcinoma. 856 38

We have developed a highly sensitive method to detect pelvic lymph node metastasis using the reverse transcriptase-polymerase chain reaction (RT-PCR) with primers specific for prostate-specific antigen (PSA) gene. Fine needle aspiration biopsy (FNAB) of pelvic lymph nodes was performed in 24 patients with prostate cancer. Each aspirated sample (0.05-0.1 ml) was divided into 2 parts; one for RNA extraction and RT-PCR to detect the fragment of PSA mRNA, and the other to smear on a slide glass for conventional cytology. The PSA gene was detected by RT-PCR in 11 FNAB samples which included not only all 6 cytologically positive and 2 cytologically class III cases but also 3 of 16 cytologically negative cases. The PSA gene was not detected by RT-PCR of FNAB samples in any of the 20 cases of bladder cancer. Thus RT-PCR for detection of the PSA gene in FNAB samples may be useful as a new diagnostic technique for detection of early lymph node metastasis in prostate cancer and an additional tool for cytological diagnosis of prostate cancer.
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PMID:[Genetic diagnosis of pelvic lymph node metastasis using fine needle aspiration samples in prostate cancer]. 895 75

We have developed a highly sensitive method to detect pelvic lymph node(LN) metastasis using reverse transcriptase-polymerase chain reaction(RT-PCR) with the primers specific for prostate-specific antigen(PSA) gene in combination with the fine needle aspiration biopsy(FNAB). The specimens were obtained from pelvic LN from 15 prostate cancer patients and 15 bladder cancer patients. The aspirated samples (0.05 approximately 0.1 ml) were used for detecting the fragment of PSA mRNA by RT-PCR and Southern blot analysis, and the rest of samples were submitted to conventional cytology. Expression of PSA gene was detected in 9 cases of FNAB samples including all 5 cytologically positive and further more 2 cytologically class III cases, and 2 of 8 cytologically negative cases. RT-PCR of FNAB samples from all cases of bladder cancer were negative for the detection of PSA gene. The sensitivity of PSA gene by RT-PCR was very high and could detect 10 degrees cancer cell. In conclusion, our study suggested that RT-PCR for detection of PSA gene in FNAB samples might become a new diagnostic tool for detection of small foci of prostatic cancer metastasis in LN and combination use of RT-PCR and cytology could greatly contribute to accuracy in diagnosis.
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PMID:[Genetic diagnosis of pelvic lymph node metastasis in prostate cancer using aspiration biopsy samples]. 899 Sep 38

Receptors for granulocyte colony-stimulating factor (G-CSFRs) have been confirmed on the cell surfaces of several non-haematopoietic cell types, including bladder cancer cells. This observation has naturally led to the hypothesis that the expression of G-CSFR on these cells may enhance their growth by G-CSF. In this study, the expression of G-CSFR was determined in both established human bladder cancer cell lines and primary bladder cancers. We studied five different human bladder cancer cell lines (KU-1, KU-7, T-24, NBT-2 and KK) and 26 newly diagnosed bladder tumours. G-CSFR mRNA expressions on cultured cell lines were determined using the reverse transcriptase polymerase chain reaction (RT-PCR) method. Furthermore, the G-CSFR binding experiments on the cultured cell lines were conducted using the Na(125)I-labelled G-CSF ligand-binding assay method. Moreover, the G-CSFR mRNA expressions on primary bladder tumour specimens were assessed using the in situ RT-PCR method. Three out of the five cultured cell lines (KU-1, NBT-2 and KK) exhibited G-CSFR mRNA signals when the RT-PCR method was used. The G-CSFR binding experiments showed an equilibrium dissociation constant (K[d]) of 490 pM for KU-1, 340 pM for NBT-2 and 103 pM for KK cells. With in situ RT-PCR, the tumour cells of 6 out of 26 primary bladder tumour specimens (23.1%) presented positive G-CSFR mRNA signals. Thus, in this study, G-CSFR expression was frequently observed on bladder cancer cells. Therefore, the clinical use of G-CSF for patients with bladder cancer should be selected with great care.
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PMID:Granulocyte colony-stimulating factor receptor expression on human transitional cell carcinoma of the bladder. 916 42

Using a reverse transcriptase-polymerase chain reaction (RT-PCR)-based quantitative analysis method, we investigated MDR1 mRNA expression levels in 58 bladder cancer specimens to determine whether MDR1 gene expression was induced or enhanced in bladder cancers during chemotherapy. In bladder cancer specimens which were obtained from patients treated with anticancer drugs, significantly higher expression levels of MDR1 mRNA were observed than in those from patients not treated with any anticancer drugs (p = 0.0134, Mann-Whitney U test). From 14 patients who had bladder cancer, clinical specimens were obtained before and after neoadjuvant intra-arterial chemotherapy. The expression levels of MDR1 mRNA were significantly higher in the post-treatment specimens than in the pre-treatment specimens (p = 0.0298, Wilcoxon signed-rank test). Of these 14 patients, 7 patients exhibited increased levels of MDR1 mRNA expression after chemotherapy. In 6 patients, there were no changes in the MDR1 mRNA expression levels before and after chemotherapy. Only one patient exhibited decreased levels of MDR1 mRNA expression after chemotherapy. No significant correlations were observed, between MDR1 mRNA expression levels and effect of the chemotherapy determined microscopically, dosage of anticancer drugs, or patient outcome. In conclusion, this study indicates that MDR1 gene expression in bladder cancers is induced and enhanced during chemotherapy. This overexpression of the MDR1 gene may contribute to resistance to anticancer drugs after repeated chemotherapy.
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PMID:[Analysis of induction of MDR1 gene expression by anticancer chemotherapy in bladder cancer]. 936 41

New, noninvasive methods for the early detection of urothelial carcinomas of the urinary bladder are needed for the diagnosis, follow-up, and screening of patients with bladder cancer. Detection of the enzyme telomerase in urine could offer these new diagnostic possibilities. The standard technique for detecting telomerase activity is the telomeric repeat amplification protocol (TRAP assay). Because of the instability of the ribonucleoprotein telomerase in an aggressive medium, such as urine, investigations conducted to date have yielded nonuniform or even contradictory findings. This study compares the detection of human telomerase RNA (hTR) by reverse transcriptase-PCR (RT-PCR) with detection of telomerase activity by the TRAP assay in the diagnosis of urothelial carcinoma of the urinary bladder. Sedimented cells obtained from urine of 30 patients with urothelial carcinoma, 15 patients with benign urological disorders, 3 patients as part of follow-up for malignant disease, and 20 healthy subjects were examined for the presence of hTR and for telomerase activity (TRAP). In patients with bladder cancer, telomerase activity was detected by the TRAP assay in only 2 of 30 specimens (7%). However, increased levels of hTR were detected by RT-PCR in 25 of the same 30 cases (83%). For patients with benign urological disorders, such as urolithiasis or urinary tract infections, hTR was detected in samples obtained from 4 of 15 patients (27%). Low hTR expression levels were found in 15% of the healthy controls. The detection of hTR by RT-PCR represents a promising new method for detecting malignant cells in urine.
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PMID:Comparison of human telomerase RNA and telomerase activity in urine for diagnosis of bladder cancer. 971 24

With the ongoing progress in human genome projects, many genes are discovered whose function and/or expression pattern are not known. Most of these genes are expressed in relatively low abundance compared to housekeeping genes such as elongation factor-1alpha and beta-actin. Gene expression is studied by Northern blot assays or by semiquantitative PCR methods. Another method is the visualization of transcripts in tissue or cell cultures by fluorescence in situ hybridization (FISH). However, for low-abundance RNA detection, this method is hampered by its limited detection sensitivity and by the interference of background signals with specific hybridization signals. Background signals are introduced by nonspecific hybridization of probe sequences or nonspecific binding of antibodies used for visualization. To eliminate background signals derived from both sources and to benefit from the peroxidase-driven tyramide signal amplification (TSA), we directly conjugated horseradish peroxidase (HRP) to oligodeoxynucleotides (ODNs) and used these probes to study in the bladder cancer cell line 5637 the expression of various cytokine genes which, according to Northern hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR) assays, are expressed at levels up to 10,000-fold less than abundantly expressed housekeeping genes. The results show that reduction of probe complexity and the limited use of immunocytochemical detection layers strongly reduces noise signals derived from nonspecific binding of nucleic acid probe and antibodies. The use of the HRP-ODNs in combination with TSA allowed detection of low-abundance cytokine mRNAs by FISH.
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PMID:Sensitive mRNA detection by fluorescence in situ hybridization using horseradish peroxidase-labeled oligodeoxynucleotides and tyramide signal amplification. 977 24

More than half of all patients with invasive urothelial cancer subsequently develop metastatic disease even after radical resection of the primary cancer. In these patients, neoplastic cells may be disseminated prior to or during the operation. A nested reverse transcriptase-polymerase chain reaction (nested RT-PCR) assay which amplifies cytokeratin (CK) 20 transcripts was used to detect cancer cells in the peripheral blood of urothelial patients. This assay was able to detect 10 bladder cancer cell line cells in a sample of ten million peripheral-blood mononuclear (PBMN) cells. CK 20-specific signals were detected in 9 (22.5%) of 40 PBMN cell samples prepared from 40 urothelial cancer patients in relation to the tumor stage, including 0/13 patients with a superficial tumor, 4/21 (19%) with a regionally invasive tumor and 5/6 (83%) with a metastatic tumor (P = 0.0002 in chi 2 test). No signals were detected in any of 25 healthy donor PBMN cell samples. The present results indicate that the CK 20 RT-PCR assay is applicable for detection of urothelial cancer cells in the peripheral blood. The assay also confirms that hematogenic dissemination occurs in invasive urothelial cancers but rarely in superficial ones.
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PMID:Detection of disseminated urothelial cancer cells in peripheral venous blood by a cytokeratin 20-specific nested reverse transcriptase-polymerase chain reaction. 1047 Feb 88

We investigated the expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP) in primary bladder cancer, its association with clinicopathologic findings, and their prognostic value. mRNA was extracted from 20 bladder cancer specimens and 6 normal bladder mucosal tissues. Relative amounts of PD-ECGF/TP mRNA were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and compared with the level of glyceraldehyde-3-phosphate dehydrogenase mRNA (used as an internal standard). PD-ECGF/TP expression was examined by immunohistochemistry in 85 patients who underwent cystectomy for bladder cancer. Serum PD-ECGF/TP levels were measured in 23 patients using a sandwich-type enzyme-linked immunosorbent assay. By RT-PCR analysis, expression of PD-ECGF/TP was found to be 7-fold higher in invasive tumors than in superficial tumors (P<0.01) and 9-fold higher than in normal bladder (P<0.01). Out of 85 transitional cell carcinoma tissue samples, 69 (81%) were evaluated as PD-ECGF/TP-positive by immunohistochemical staining. PD-ECGF/TP expression correlated significantly with tumor grade (P = 0.001), depth of invasion (P = 0.012), and lymphatic invasion (P = 0.01). No correlation was found between expression of PD-ECGF/TP and the number of tumors, tumor configuration, lymph node involvement, venous invasion, c-erbB-2 expression, or overall survival. We could not detect a significant serum level of PD-ECGF/TP in any patient. The results suggest that PD-ECGF/TP might give valuable information for bladder cancer management, though it may not be a good new tumor marker for bladder cancer.
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PMID:Expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase in human bladder cancer. 1066 52

Cytogenetic and loss of heterozygosity (LOH) studies demonstrated chromosome 3p deletions in transitional cell carcinoma (TCC). We recently cloned the tumor suppressor gene FHIT (fragile histidine triad) at 3p14.2, one of the most frequently deleted chromosomal regions in TCC of the bladder, and showed that it is the target of environmental carcinogens. Abnormalities at the FHIT locus have been found in tumors of the lung, breast, cervix, head and neck, stomach, pancreas, and clear cell carcinoma of the kidney. We examined six TCC derived cell lines (SW780, T24, Hs228T, CRL7930, CRL7833, and HTB9) and 30 primary TCC of the bladder for the integrity of the FHIT transcript, using reverse transcriptase-polymerase chain reaction (RT-PCR) to investigate a potential role of the FHIT gene in TCC of the bladder. In addition, we tested expression of the Fhit protein in the six TCC-derived cell lines by Western blot analysis and in 85 specimens of primary TCCs by immunohistochemistry. Three of the six cell lines (50%) did not show the wild-type FHIT transcript, and Fhit protein was not detected in four of the six cell lines (67%) tested. Fhit expression also was correlated with pathological and clinical status. A significant correlation was observed between reduced Fhit expression and advanced stage of the tumors. Overall, 26 of 30 (87%) primary TCCs showed abnormal transcripts. Fhit protein was absent or greatly reduced in 61% of the TCCs analyzed by immunohistochemistry. These results suggested that loss of Fhit expression may be as important in the development of bladder cancer as it is for other neoplasms caused by environmental carcinogens.
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PMID:Loss of FHIT expression in transitional cell carcinoma of the urinary bladder. 1066 70


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