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)

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

The aim of this study was to determine the presence of haematogenous neoplastic cells in patients with prostate cancer. Circulating prostate cells can be detected in cancer patients by using a nested-reverse transcriptase-polymerase chain reaction assay (RT-PCR), for prostate-specific membrane (PSM) antigen mRNA. This sensitive nested RT-PCR assay may play a crucial role in the administration of adjuvant therapy of patients with prostate adenocarcinoma.
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PMID:The use of nested RT-PCR of prostate-specific membrane antigen in blood cells: implications for the detection of haematogenous neoplastic cells in patients with prostate adenocarcinoma. 984 60

Objectives: We investigated modulation of cell growth and prostate-specific antigen (PSA) gene expression in prostatic cancer cells by the luteinizing hormone-releasing hormone analog (LH-RHa), leuprorelin acetate, alone or combined with other agents. Methods: The effect of the analog on proliferation of both androgen-sensitive and -insensitive prostate cancer cells, maintained in different culture conditions, was evaluated by cell counts at various intervals of time. Basal expression of PSA gene and its variations were determined by a reverse transcriptase-polymerase chain reaction assay. Results: LH-RHa is ineffective in regulating cell growth, when used alone in both hormone-sensitive and -insensitive cell lines. Nevertheless, it counteracts the stimulatory action of androgens on proliferation of LNCaP cells, which respond to low concentrations of dihydrotestosterone. Moreover, LH-RHa has an inhibitory effect on the mitogenic action of epidermal growth factor (EGF) in androgen-unresponsive PC-3 cells. The analog reduces PSA gene expression in both hormone-sensitive and -insensitive cells. Interestingly, it counteracts the gene expression induced by androgens in LNCaP cells and by EGF in PC-3 cells. Conclusions: These data show that LH-RHa may behave like a negative growth factor, which directly regulates cell growth and PSA gene expression. Moreover, our findings support the idea that growth factors may interfere with the androgen signalling pathway.
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PMID:Effect of Leuprorelin Acetate on Cell Growth and Prostate-Specific Antigen Gene Expression in Human Prostatic Cancer Cells. 985 46

The potential use of prostate secretory protein of 94 amino acids (PSP94) as a diagnostic biomarker or a therapeutic agent for prostate cancer has been reported. In order to establish an animal model to further elucidate on its biological role, we cloned the mouse PSP94 cDNA (approximately 500 bp) by reverse transcriptase-polymerase chain reaction (RT-PCR) and disclosed its genomic structure. The whole mouse PSP94 gene (approximately 23 kb) was amplified by long and accurate-PCR and also cloned by screening of a mouse embryo stem-cell genomic library. Computational and statistical analyses have demonstrated several highly conserved characteristics of PSP94 among different species. Comparison of PSP94 from human, two primates, pig, and rodents revealed that the most significant feature is that PSP94 is rich in cysteines (10% of the total sequence) and their positions are highly conserved. The three intron-four exon structure of the human PSP94 gene and the consensus sequence (....GT-intron-AG...) for mRNA splicing are also strongly conserved. A high divergence in cDNA sequence in the protein-coding region and also in the genomic sequence of PSP94 was also observed among these species. Comparing with alpha-globin, a typical evolutionally conserved gene, with the PSP94 gene, the rate of nonsynonymous changes per site per year (kN) is 2 to 6 times higher, indicating that PSP94 gene has been under far fewer evolutionary constraints than other genes and has a potential role as a species barrier in reproductive biology. In order to test this hypothesis, we investigated the gene expression of PSP94 and its tissue distribution in various rodent tissues by RT-PCR and in situ hybridization (ISH). Gene expression was found only in the prostate, suggesting that PSP94 is probably more tissue specific in the prostate of rodents than in mammals. The ISH analysis also revealed a prostate lobe-specific expression of the PSP94 gene in both mice and rats. It was strongly expressed in the lateral prostate, but the findings were negative in the dorsal and ventral lobe. Therefore, it is hypothesized that one of the primary functions of rodent PSP94, as a major prostate secretory protein, is related to reproductive biology.
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PMID:cDNA, genomic cloning, and gene expression analysis of mouse PSP94 (prostate secretory protein of 94 amino acids). 1002 5

The reverse transcriptase-polymerase chain reaction (RT-PCR) assay is an extremely sensitive technique for the detection of circulating cells expressing prostate-specific antigen (PSA) in prostate cancer patients. This article reviews the literature on the use of this technique as a preoperative parameter to predict both extraprostatic disease and PSA recurrence after radical prostatectomy. Despite the relative consensus regarding the increase in RT-PCR-positivity with tumor stage (i.e., clinically localized vs metastatic prostate cancer), the use of RT-PCR as a clinical staging modality is controversial. To date, more than 16 institutions have evaluated the RT-PCR test in prostate cancer. Of these institutions, only two have reported the utility of RT-PCR as a staging modality and three have reported the utility of the test in predicting PSA recurrence. Before further conclusions are drawn regarding the clinical utility of RT-PCR in prostate cancer patients and its routine use is advocated, a larger patient population needs to be studied and followed for longer periods.
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PMID:Molecular staging of prostate cancer: dream or reality? 1007 69

The detection of prostate-specific membrane antigen (PSM) mRNA in the peripheral blood of prostate cancer patients by a nested reverse transcriptase-polymerase chain reaction (RT-PCR) assay is a useful and sensitive method for the identification of small foci of metastatic lesions. In this study, a nested RT-PCR assay was performed using the two different PSM-derived oligonucleotide primer sets reported by Israeli et al. and Loric et al. (termed PSM primers-1 and primers-2, respectively, in this report), and the differences in the specificity and sensitivity of these primer sets for detecting prostate cancer cells in the blood are discussed. The PCR assay using PSM primers-1 showed DNA bands for 4 of 7 cases of metastatic prostate cancer and amplified the untreated genomic DNA, while that using PSM primers-2 showed 6 bands without the amplification of the genomic DNA. In conclusion, PSM primers-2 is superior to PSM primers-1 for the detection of PSM mRNA in the peripheral blood of prostate cancer patients.
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PMID:Prostate-specific membrane antigen-derived primers in a nested reverse transcription polymerase chain reaction for detecting prostatic cancer cells. 1018 95

To determine the potential risk of hematogenous dissemination of prostate cancer cells during radical prostatectomy (RP), we investigated the pre- and intraoperative circulating prostate-specific antigen (PSA) mRNA in patients with clinically localized prostate cancer, with special reference to neoadjuvant hormonal therapy (NHT). Using a nested reverse transcriptase (RT) polymerase reaction (PCR) assay, PSA mRNA in the peripheral blood was evaluated pre- and postoperatively in a total of 23 patients, 10 of whom received NHT with antiandrogens. The RT-PCR assay employed detected one LNCaP cell in 10(7) mononuclear blood cells, and showed no positive signal in the blood samples from all 15 healthy controls. Pre- and intraoperative circulating PSA mRNA was positive in 11 (48%) and 18 patients (78%), respectively. All 11 patients with positive preoperative PSA mRNA continued to be positive during RP, and seven (58%) of 12 patients with negative preoperative PSA mRNA had a positive conversion. Although the patients' ages, preoperative serum PSA values and clinical or pathological stages were not associated with the pre- and intraoperative PSA mRNA results, the NHT group showed a significantly lower incidence of preoperative PSA mRNA positivity (2/10) than the group receiving RP alone (9/13) (20% vs 69%, P = 0.036). NHT, however, showed no suppressive effect on either intraoperative positivity or positive conversion of circulating PSA mRNA. The present study suggests that a substantial number of patients receiving RP are at risk of hematogenous dissemination, and NHT with antiandrogens has a minimal or no suppressive effect on the circulating PSA mRNA during surgical manipulation of the prostate. Because the clinical significance of circulating cancer cells remains to be determined, long-term follow-up in association with the circulating cancer cells assessed by the RT-PCR is essential in order to establish the role of molecular staging as well as NHT.
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PMID:Circulating prostate-specific antigen mRNA during radical prostatectomy in patients with localized prostate cancer: with special reference to neoadjuvant hormonal therapy. 1046 Sep 2

Background: Preoperative staging for prostate cancer underestimates the final pathology stage in approximately 40-50% of the cases. Previous work from our institution demonstrated that an enhanced reverse transcriptase polymerase chain reaction (RT-PCR) assay for prostate-specific antigen (PSA) enabled more accurate staging of presumably localized prostate cancer. The goal of the current study is to determine if needle biopsy results when combined with the RT-PCR for PSA assay are a better predictor of final pathology stage. Methods and Results: One hundred sixty-two men with needle biopsy-diagnosed prostate cancer had blood drawn for the RT-PCR for PSA assay before undergoing radical prostatectomy. Polymerase chain reaction primers specific for the PSA gene were run, along with appropriate controls. Tumor was characterized using the TMN staging system: organ confined (pT2a-c), capsular penetration (pT2a-b), seminal vesicle involvement (pT3c). Surgical margins and lymph nodes were also evaluated. Of the 162 patients, the majority had localized disease by digital rectal examination: T2 = 97%, and T3 = 3%. On needle biopsy, 48 cases (30%) had a Gleason score >/=7 and 35 cases (22%) had perineural involvement (PNI). The RT-PCR for PSA assay was positive in 50 patients (31%). Final pathology revealed 39% of patients had pT3 disease; none of the 162 patients had lymph node involvement. Statistical analysis revealed that a Gleason score >/=7 had 81% specificity and 46% sensitivity in predicting pT3 disease (odds ratio 3.6). The presence of PNI on needle biopsy was 89% specific and 38% sensitive in predicting pT3 disease (odds ratio, 4.9). The RT-PCR for PSA assay was 89% specific and 62% sensitive in predicting pT3 disease (odds ratio, 13.0). All 14 cases with both RT-PCR for PSA and PNI positivity had pT3 disease. Logistic regression analysis demonstrated the independent predictive strength of PNI on needle biopsy, Gleason score >/=7, and RT-PCR for PSA positivity for identifying pT3 disease; their combined odds ratio was more than 180. Conclusions: Using the RT-PCR for PSA assay in conjunction with needle biopsy results increases the predictive strength for pT3 disease in patients with presumed organ-confined prostate carcinoma.
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PMID:Enhanced Reverse Transcriptase Polymerase Chain Reaction for Prostate-specific Antigen Combined With Needle Biopsy Results: A Superior Predictor of pT3 Disease. 1046 1

The activity and expression of transgene beta-galactosidase (lacZ) by replication-deficient adenoviral vectors (Ad-lacZ) containing prostate-specific promoters were compared using an in vivo canine model. The prostate tissue-specific promoters were prostate-specific antigen, probasin, and mouse mammary tumor virus long-terminal repeat, which were fused separately to an Escherichia coli lacZ gene. Dogs underwent laparotomy, and adenoviral vectors were delivered by direct intraprostatic injection. At 72 hours postinjection, the prostate and various other organs were harvested to evaluate the degree of prostate expression and dissemination of adenoviral vectors. Expression of lacZ in tissues was determined by 5-bromo-4-chloro-3-indolyl beta-D-galactoside staining, beta-galactosidase assay, and E. coli lacZ reverse transcriptase-polymerase chain reaction (PCR). The presence of adenoviral DNA sequences in canine tissues was determined by PCR using primers specific for the type 5 adenoviral genome. All three of the prostate-specific adenoviruses tested effectively expressed the lacZ gene in the canine prostate, but expression levels were lower than that of the control viral vector AdRSVlacZ following intraprostatic injection. By PCR, adenoviral vector DNA was detected in other organs and tissues, including the bladder and vas deferens. However, reverse transcriptase-PCR analysis revealed that prostate-specific Ad-lacZ vectors only transcribed lacZ mRNA in the prostate and not in nonprostatic tissues. Thus, these novel prostate-specific adenoviral vectors each have equal in vivo expression exclusively in the prostate and may potentially be used for prostate cancer gene therapy.
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PMID:In vivo expression of prostate-specific adenoviral vectors in a canine model. 1050 56

An important biological feature of prostate cancer (PCa) is its marked preference for bone marrow as a metastatic site. To identify factors that may support the growth of PCa in bone marrow, expression of receptor and nonreceptor tyrosine kinases by androgen-independent PCa bone marrow metastases was assessed. Bone marrow biopsies largely replaced by PCa were analyzed using reverse transcriptase-polymerase chain reaction amplification with degenerate primers that amplified the conserved kinase domain. Sequence analyses of the cloned products demonstrated expression of multiple kinases. Expression of the receptor and nonreceptor tyrosine kinases, alpha platelet-derived growth factor receptor and Jak 1, respectively, was confirmed by immunohistochemistry. In contrast, the type 1 insulin-like growth factor receptor, thought to play a role in PCa development, was lost in metastatic PCa. These results implicate several specific growth factors and signaling pathways in metastatic androgen-independent PCa and indicate that loss of the type 1 insulin-like growth factor receptor contributes to PCa progression.
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PMID:Tyrosine kinases expressed in vivo by human prostate cancer bone marrow metastases and loss of the type 1 insulin-like growth factor receptor. 1051 9


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