We, however, lack the description of the expression of these molecules in human prostate cancer PCC and in benign prostate hyperplasia BPH. Results CB1 was identiWed in epithelial and smooth muscle cells types of the human prostate, whereas TRPV1 was exclusively localized to the mucosal cells.
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Czifra · K. Nyeste · R. Marincsák · B. Tóth · L. Kovács · T. Keywords Benign prostate hyperplasia · Cannabinoid receptor-1 CB1 · Human prostate bph vs prostate cancer histology · Transient receptor potential vanilloid-1 TRPV1 Introduction Although the metabotropic, G-protein coupled cannabinoid receptor-1 CB1 and the ligand-gated, calcium-permeable ion channel transient receptor potential vanilloid subtype-1 TRPV1 possess markedly distinct structural properties, they share a wide array of common features both in localization and function.
Namely, these receptors were originally described on speciWc neuronal structures and were implicated in the regulation of behavior, learning, and memory for CB1and in pain and temperature sensation for TRPV1 Caterina and Julius ; De Petrocellis et al.
In addition, they can be activated by numerous exogenous e. Moreover, it was also postulated that the CB1- and TRPV1-coupled signaling mechanisms very often interact, hence resulting in a complex, bi-directional regulatory relationship Caterina and Julius ; De Petrocellis et al. Of great importance, other studies also revealed that CB1 and TRPV1 are additionally expressed on numerous non-neuronal cell types such as, e.
On these cells, it was suggested that the two receptors and their endogenous ligands play key roles in the regulation of such processes as, e.
The functional role of the cannabinoid and vanilloid systems in the regulation of cell proliferation and death was also described in human prostate cancer PCC derived cell lines.
Namely, it was shown that exogenous or endogenous cannabinoid and vanilloid ligands induced in vitro growthinhibition and apoptotic or necrotic cell death of the PCC cell lines expressing CB1 and TRPV1 Nithipatikom et al.
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Furthermore, recent reports presented that vanilloids in vivo eVectively prevented the growth or induced the regression of xenograft tumors induced by PCC-derived cell lines Sanchez et al. However, although the in situ expression of both receptors was described in healthy prostate tissue and, for TRPV1, in samples from benign prostate hyperplasia [BPH] patients Galiegue et al.
The study involved 35 cases of PCC 8 cases of G1, 10 cases of G2, 8 cases of G3, and 9 cases of G4 grades removed by prostatectomy, 12 samples of BPH removed by transurethral resection, and 9 samples of normal healthy bph vs prostate cancer histology obtained from transplantation donors.
In each case, the tissues were divided into three parts according to our previous report Varga et al. In addition experiments, to further assess speciWcity of the immunostaining, primary labeling was performed using goat C-terminus-speciWc antibodies: anti-CB1 K, sc, dilution and anti-TRPV1 D, sc, dilution.
The application of these latter primary antibodies resulted in identical staining patterns data not shown. For negative controls of the labeling procedure, antibodies were pre-absorbed by control blocking peptides provided along with appropriate protocols by the manufacturer Santa Cruz.
For positive controls, human organ-cultured hair follicle and full-thickness skin sections according to our previous reports, Bodó et al. When double Xuorescence labeling of CB1 was performed Bodó et al. Subsequently, tissues were incubated with species-matched i.
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HRP-conjugated secondary antibodiesBioRad were then employed, and the immunoreactive bands were visualized by enhanced chemiluminescence Pierce. We, however, also found that while the TRPV1-ir was clearly restricted to the epithelial cells, the CB1-speciWc signals besides the previously described epithelial cells Ruiz-Llorente et al.
To identify those cells types that express CB1, double Xuorescence immunolabeling was performed. As seen in Fig. We then started to investigate the expressions of the two receptors in various hyperproliferative diseases of the human prostate.
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However, we observed a dramatically increased expression of CB1, both in the epithelial and smooth muscle cells in BPH Figs. Similarly, we found that the level of CB1 was also increased in samples from PCC patients, especially in the epithelial cells Figs.
Although the above immunohistochemical determination strongly suggested that the level of CB1 increased both in BPH and PCC, whereas that of TRPV1 elevated in PCC the semi-quantitative nature of this technique did not permit exact, quantitative determination of the expressions of the molecules. In addition, to measure a putative relationship between the expressions of the receptors Fig. CB1 and TRPV1 immunoreactivity on healthy prostate control and diseased prostate tissues, as revealed by alkaline phosphatase method.
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Negative control was obtained by pre-absorbing the antibodies with appropriate blocking peptides. Interestingly, however, no correlation was observed between the degree of CB1 expression and the increasing tumor grades. Namely, when values of the groups of various tumor grades were statistically compared to one another, the only signiWcant diVerence was found between the G1 and G2 PCC groups, but only when measured by immunoblotting Fig.
Similarly, the expression of the receptor was not altered in the G1 low malignancy PCC samples. In addition, the statistical analysis showed that, as opposed to Wndings with CB1, the expression of TRPV1 both the protein and mRNA levels in almost all cases increased in parallel and gradually with the degree Ki gyakrabban beteg prosztata rák malignancy, i.
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Discussion The novel results presented in this paper provide the Wrst evidence that CB1 is signiWcantly elevated in human PCC tissues mostly in the epithelial elements when compared to normal healthy prostate. Therefore, although further studies e. The amount of CB1 b and TRPV1 d protein levels in a given tissue was quantitated by densitometry and normalized to the optical density values of cytochrome C of the same sample.
Normalized values were then averaged within the same healthy, diseased group. In all panels, points represent the mean § SEM of normalized values of numerous samples in each group.
These data may suggest that in contrast to human hepatocellular carcinoma where the expression of CB1 well correlated with the clinical outcome of the disease Xu et al. There were, however, two important diVerences when compared the expression patterns of the two receptors.
Interestingly, previous studies on another human urogenital tract tumor, i. Namely, Lazzeri et al. The explanation of this contradiction is unknown yet, for the potential therapeutic targeting of TRPV1 to be deWnitely determined.
The authors state no competing Wnancial interest. J Bph vs prostate cancer histology Dermatol — Annu Rev Neurosci — Br J Pharmacol — Nat Rev Drug Discov — Oncogene — Eur J Biochem — J Mol Med — Eur Urol — Cancer Res — Handb Exp Pharmacol — J Clin Invest — Prostate — Eur J Pharmacol — Apoptosis — J Urol — Cancer Genet Cytogenet — Endocr Relat Cancer —