-
[show abstract]
[hide abstract]
ABSTRACT: To date, the calcium-regulated membrane guanylate cyclase Rod Outer Segment Guanylate Cyclase type 1 (ROS-GC1) transduction
system in addition to photoreceptors is known to be expressed in three other types of neuronal cells: in the pinealocytes,
mitral cells of the olfactory bulb and the gustatory epithelium of tongue. Very recent studies from our laboratory show that
expression of ROS-GC1 is not restricted to the neuronal cells; the male gonads and the spermatozoa also express ROS-GC1. In
this presentation, the authors review the existing information on the localization and function of guanylate cyclase with
special emphasis on Ca2+-modulated membrane guanylate cyclase, ROS-GC1, in the testes. The role of ROS-GC1 and its Ca2+-sensing modulators in the processes of spermatogenesis and fertilization are discussed.
Molecular and Cellular Biochemistry 04/2012; 334(1):169-179. · 2.06 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Metastasis is a common feature of many advanced stage cancers and metastatic spread is thought to be responsible for cancer progression. Most cancer cells are localized in the primary tumor and only a small population of circulating tumor cells (CTC) has metastatic potential. CTC amount reflects the aggressiveness of tumors, therefore their detection can be used to determine the prognosis and treatment of cancer patients.The aim of this study was to evaluate human chorionic gonadotropin beta subunit (CGB) and gonadoliberin type 1 (GNRH1) expression as markers of tumor cells circulating in peripheral blood of gynecological cancer patients, indicating the metastatic spread of tumor.
CGB and GNRH1 expression level in tumor tissue and blood of cancer patients was assessed by real-time RT-PCR. The data was analyzed using the Mann-Whitney U and Spearman tests. In order to distinguish populations with homogeneous genes' expression the maximal likelihood method for one- and multiplied normal distribution was used.
Real time RT-PCR results revealed CGB and GNRH1 genes activity in both tumor tissue and blood of gynecological cancers patients. While the expression of both genes characterized all examined tumor tissues, in case of blood analysis, the transcripts of GNRH1 were found in all cancer patients while CGB were present in 93% of patients. CGB and GNRH1 activity was detected also in control group, which consisted of tissue lacking cancerous changes and blood of healthy volunteers. The log-transformation of raw data fitted to multiplied normal distribution model showed that CGB and GNRH1 expression is heterogeneous and more than one population can be distinguished within defined groups.Based on CGB gene activity a critical value indicating the presence of cancer cells in studied blood was distinguished. In case of GNRH1 this value was not established since the results of the gene expression in blood of cancer patients and healthy volunteers were overlapping. However one subpopulation consists of cancer patient with much higher GNRH1 expression than in control group was found.
Assessment of CGB and GNRH1 expression level in cancer patients' blood may be useful for indicating metastatic spread of tumor cells.
Journal of Translational Medicine 08/2011; 9:130. · 3.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To date, the calcium-regulated membrane guanylate cyclase Rod Outer Segment Guanylate Cyclase type 1 (ROS-GC1) transduction system in addition to photoreceptors is known to be expressed in three other types of neuronal cells: in the pinealocytes, mitral cells of the olfactory bulb and the gustatory epithelium of tongue. Very recent studies from our laboratory show that expression of ROS-GC1 is not restricted to the neuronal cells; the male gonads and the spermatozoa also express ROS-GC1. In this presentation, the authors review the existing information on the localization and function of guanylate cyclase with special emphasis on Ca(2+)-modulated membrane guanylate cyclase, ROS-GC1, in the testes. The role of ROS-GC1 and its Ca(2+)-sensing modulators in the processes of spermatogenesis and fertilization are discussed.
Molecular and Cellular Biochemistry 11/2009; 334(1-2):169-79. · 2.06 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The survivin is the protein involved in regulation of basic and cycle-specific functions of cells both in normal and cancer tissue. Recent studies present survivin as a factor having the leading role in the regulation of apoptosis and mitosis as well as a target of anticancer therapy. The employing of survivin in this therapy is based on its high expression level in most human cancers, as well as its association with the disease's progression. The aim of our study was to evaluate the expression and localization of survivin's gene product on the protein level in different types of pituitary tumors and normal pituitary. The coexpression of survivin and proliferating cell nuclear antigen - PCNA in pituitary was also examined.
The study was conducted on the postoperative pituitary tumors tissue taken during standard neurosurgical removal of tumor from 43 patients. The group of patients consists of 23 women and 20 men, aged from 27 to 71 years. As a control of the study three normal pituitary tissues obtained at the autopsy were used. Evaluation of survivin and PCNA expression was performed using immunohistochemical staining.
The study demonstrated the presence of survivin in all analyzed by us pituitary tumors. Survivin was present also in normal pituitary tissue. The protein was localized mainly in cell's nuclei, however the less intense immunostaining was observed also in the cytoplasm of pituitary tumors cells. Furthermore survivin was found in normal pituitary, but the positive immunostaining was limited to a single cells. The analysis of pituitary tumor cells proliferation index based on PCNA reactivity showed that survivin is coexpressed with PCNA, especially in invasive tumors.
The study documented the presence of survivin in different types of pituitary tumors as well as in normal pituitary. Additionally the coexpression of survivin and PCNA in tumor cells was shown. The expression of survivin in both normal and cancer pituitary cells suggests that it may play an important role in regulation of the gland's proliferation.
Neuro endocrinology letters 10/2009; 30(4):477-81. · 1.30 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Evaluation of the presence of a Ca(2+)-regulated membrane guanylate cyclase signal transudation system in the spermatozoa.
Experimental study.
Research university laboratory.
Human sperm obtained from healthy donors who met the criteria of the World Health Organization for normozoospermia and bovine semen collected from bulls of proven fertility.
Radioimmunoassay and immunohistochemistry of human and bovine spermatozoa.
The membrane guanylate cyclase activity and the presence of membrane guanylate cyclase transduction machinery components in the spermatozoa.
The identity of a Ca(2+)-modulated membrane guanylate cyclase transduction machinery in human and bovine spermatozoa has been documented. The machinery is both inhibited and stimulated within nanomolar to semimicromolar range of free Ca(2+). The transduction component of this machinery is the rod outer segment membrane guanylate cyclase type 1 (ROS-GC1). The enzyme coexists with three Ca(2+)-dependent modulators: guanylate cyclase activating protein type 1 (GCAP1), S100B and neurocalcin delta. ROS-GC1 and its modulators are present in the heads and tails of both species' spermatozoa.
The coexpression of ROS-GC1 and its activators in spermatozoa suggests that the Ca(2+)-modulated ROS-GC1 transduction system may be a part of the fertilization machinery.
Fertility and sterility 01/2009; 93(3):904-12. · 3.97 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: It has been suggested that ghrelin synthesized locally in pituitary regulates the function and growth of pituitary cells in autocrine/paracrine way and might be an important factor of pituitary tumorogenesis. The expression of ghrelin receptor in neoplastic cells of pituitary adenomas has also been demonstrated. In vitro studies confirmed that ghrelin stimulates the proliferation of somatotroph cells in GH3 cell line. The presence of both ghrelin mRNA and protein was detected in a number of benign and malignant neoplasms as well as in neoplastic cells of the tissues which do not express ghrelin in physiological conditions. This study showed the presence of ghrelin mRNA and its protein in different types of pituitary adenomas.
The samples of 37 pituitary adenomas were obtained during standard neurosurgical tumor removal. The study tissues included 20 somatotroph tumors (15 patients treated and 5 patients untreated with octreotide LAR before the surgery), 12 nonfunctioning adenomas, 4 prolactinomas and 1 ACTH-secreting tumor. The control included samples of normal mucous membrane of the stomach and normal pituitaries. Expression of ghrelin mRNA was studied in 28 pituitary adenomas by RT-PCR. Immunohistochemical evaluation of ghrelin presence was performed in 34 tumors.
The presence of ghrelin gene transcripts was demonstrated in 10 out of 15 examined somatotroph tumors (obtained from patients treated with octreotide LAR before the surgery) and also in 2 out of 4 samples of prolactinomas, 7 out of 8 of nonfunctioning tumors and in 2 samples of normal pituitary. Immunohistochemical analyses revealed the presence of the protein in all 5 examined somatotroph tumors obtained from patients not treated prior to the surgery and in 10 out of 15 tumors obtained from patients treated with octreotide LAR. The peptide was detected also in 10 out of 12 examined nonfunctioning tumors and in 2 examined PRL-secreting tumors. The immunostaining for ghrelin was not shown in normal pituitaries.
The study demonstrated that ghrelin gene is expressed in somatotroph adenomas, both treated and untreated with octreotide LAR before the surgery, and also in other types of pituitary adenomas (prolactinomas and nonfunctioning adenomas).
Neuro endocrinology letters 01/2009; 29(6):929-38. · 1.30 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Survivin is a member of the inhibitor of apoptosis protein family, which was recently showed to be expressed by different benign and malignant human tumors. Still very little is known about survivin expression in pituitary tumors. In spite of the fact that pituitary tumors in histological examination are usually benign, in the clinical process a certain number of pituitary adenomas is capable of aggressive growth, recurrence and invasion of the surrounding structures. The aim of the present study was to assess the presence of survivin transcripts and protein in different types of pituitary tumors and to evaluate survivin expression levels in invasive and non-invasive pituitary tumors.
The analyzed material consisted of tumor tissue samples obtained during standard neurosurgical removal of the tumor from 23 patients in whom acromegaly (n=14), non-functioning pituitary tumor (n=6), prolactinoma (n=2) and corticotropinoma (n=1) were diagnosed. As a control of the study normal pituitary tissue obtained at autopsy was used. Amplification of survivin gene using sequence specific primers and qRT-PCR method and immunohistochemical staining with primary polyclonal antibodies against human survivin were performed.
Our study demonstrated the presence of survivin mRNA in all 23 analyzed pituitary tumors. Survivin expression was also observed in normal pituitary, but the level of its expression was 6-fold lower than in tumors tissue when studied by real time RT-PCR. The difference between the levels of survivin expression in invasive and non-invasive tumors was not statistically significant. Immunohistochemical analyses revealed the presence of the protein in both normal and tumor tissue of pituitary. Immunostaining of tumor tissue was not uniform. Survivin was observed mainly in the nuclei of cells collected in clusters. The presence of the protein in normal pituitary was restricted to small population of cells.
The present study showed that overexpression of survivin is characteristic for pituitary tumors. Further analysis of this protein expression profile should demonstrate whether survivin might be use as a prognostic marker in diagnosis and therapy of pituitary adenomas.
Neuro endocrinology letters 01/2009; 29(6):1033-7. · 1.30 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The aim of the study was to test the effect of photodynamic therapy on fibroblast proliferation in vitro using protoporphyrin IX as a photosensitizer.
The abnormal proliferation of synovial tissue serves as a propagator of immune response and tissue damage in rheumatoid arthritis. Since the synovial fibroblasts mediate most relevant pathways of joint destruction, they constitute an important target for novel therapeutic approaches. Photodynamic therapy, which is the clinically applied treatment for various tumors, as well as for some non-oncologic diseases, is based on administration of an exogenous photosensitizer which is used to induce cell death of fibroblast cells.
The photosensitizing effects of protoporphyrin IX (PpIX) were studied in the 3T3 cell line that served as a model of fibroblasts and HeLa cells cultured in vitro.
The number of fibroblasts and HeLa cells treated with photo-activated PpIX decreased significantly. The compound affects the viability of study cells, causing necrosis of 3T3 cells and apoptosis of HeLa cells.
The light-activated protoporphyrin affected proliferation of both 3T3 and HeLa cells. No effects of the phototherapy were seen in the form of apoptosis of 3T3 cells, whereas the induction of cell death in HeLa cells was detected.
Photomedicine and laser surgery 08/2008; 26(4):343-7. · 1.76 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Secretion of human chorionic gonadotropin, especially its beta subunit by malignant trophoblastic tumors and varieties of tumors of different origin is now well documented; however the role of hCG in tumorogenesis is still unknown.
This study documents the molecular presence of human chorionic gonadotropin beta subunit in uterine cervix cancer tissues and investigates a novel technique to reduce hCGbeta levels based on expression of a modified U1 snRNA as a method to study the hormone's role in biology of human cervical cancer cells cultured in vitro. The property of U1 snRNA to block the accumulation of specific RNA transcript when it binds to its donor sequence within the 3' terminal exon was used. The first 10 nucleotides of the human U1 snRNA gene, which normally binds to the 5'ss in pre-mRNA were replaced by a sequence complementary to a 10-nt segment in the terminal exon of the hCGbeta mRNA. Three different 5' end-mutated U1 snRNA expression plasmids were tested, each targeting a different sequence in the hCGbeta mRNA, and we found each one blocked the expression of hCGbeta in HeLa cells, a cervix carcinoma cell line, as shown by immunohistochemistry and qRT-PCR. Reduction of hCGbeta levels resulted in a significantly increased apoptosis rate with almost 90% of cells transfected with modified anti-hCGbeta U1 snRNAs showing morphological changes characteristic of the apoptotic process.
These data suggest that human chorionic gonadotropin beta subunit may act as a tumor growth-stimulating factor.
Molecular Cancer 02/2008; 7:26. · 3.99 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The research on the expression and mutations of DAZ and its homologues in human and other mammals suggests that protein products of these genes can mainly affect development of germinal cells. The aim of the present study was to analyze the expression of the DAZ gene in seminiferous tubules of six men with spermatogenic disorders (hypospermatogenesis and spermatogenic arrest). The results based on the RT-PCR IS technique demonstrated that the DAZ product was present only in some seminiferous tubules and the fluorescence intensity was different within individual tubules. The most intense fluorescence characterised the spermatogonia, especially these organised in small groups inside separate tubules. In the patients with spermatogenic arrest at the spermatocyte stage, the DAZ gene transcripts were also found in primary spermatocytes. However, the fluorescence intensity of primary spermatocytes, except the fluorescence of the spermatocytes localised upon the lumen, was weaker than the fluorescence of spermatogonia. The results of our study showed that DAZ gene activity seems to correspond to the proliferative activity of stem cells of germinal epithelium.
Folia Histochemica et Cytobiologica 02/2006; 44(2):123-6. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Survivin has received great attention due to its expression in many human tumours and its potential as a therapeutic target in cancer. Its expression is developmentally regulated: present during fetal development, it is undetectable in terminally differentiated normal adult tissue. Survivin expression has been described to be cell cycle-dependent and restricted to the G2-M checkpoint, where it inhibits apoptosis in proliferating cells.
The aim of our study was to determine the survivin expression in different types of pituitary adenomas.
Tissue samples were obtained during surgical removal of the tumour from 12 patients with diagnosed: acromegaly in seven cases, non-functioning pituitary tumours in four cases and prolactinoma in one case. Six patients with acromegaly received long-acting somatostatin analogues before tumour resection. After RNA extraction and cDNA synthesis, the amplification of specific survivin's gene fragment was performed.
In agreement with the current view that survivin is a tumor-associated antigen, highly expressed in various tumours, we found the presence of survivin expression as a characteristic feature of human pituitary adenomas. The findings of our study demonstrated the presence of an active survivin gene in all twelve analysed pituitary tumours.
Based on these findings, we conclude that the estimation of survivin expression in human pituitary tumours may help predict tumour growth and prognosis.
Neuro endocrinology letters 07/2005; 26(3):209-12. · 1.30 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The ovary is a tissue, which is sensitive to gonadotropins. Gonadotropins receptors are present in the epithelium and stroma of the ovary. Data from epidemiological study clearly shows, that development of tumor is correlated with morphological and biochemical processes taking place in the gonad. The role of gonadotropins, as well as the role of their receptors in the proliferation of the ovarian tissue was shown. Further research is needed to explain the molecular mechanism of gonadotropin activity in ovarian tissue, however the important role of gonadotropins in the pathology of ovarian tissue is already well established.
Ginekologia polska 03/2005; 76(2):153-62. · 0.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Recent studies demonstrated that besides placenta and malignant trophoblastic tumors, hCG and especially its beta-subunit is secreted by a varieties of tumors of different origin. The aim of the present investigation was to determine the expression pattern of human chorionic gonadotropin gene in ovarian cancer tissue. The study included 8 patients with epithelial ovarian carcinoma. The expression and distribution of hCGbeta mRNA was assessed by in situ RT-PCR method. The semi-quantitative assessment was performed using computer image analysis. Transformation of the images into the pseudocolour scale showed a clear difference in fluorescence intensity among individual cancer cells. The intensity of ISRT-PCR products corresponding with expression level of hCGbeta demonstrated that its production by individual cancer cells is different. In all studied specimens of the ovarian carcinoma tissue, cancer cells characterized by the presence of active hCGbeta gene were found, whereas noncancerous tissue demonstrated lack of the gene expression. Thus, the study clearly shows that the expression of hCGbeta is the feature of ovarian cancer tissue.
Folia Histochemica et Cytobiologica 02/2004; 42(2):123-6. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Apoptosis of endocrine cells represents one of factors significantly affected the control of hormones secretion. Present study aimed to examine whether treatment of somatotropinoma types tumours with somatostatine analogue (lanreotide) results in apoptosis.
The studies were performed on 35 patients with somatotropinoma type tumours on whom adenomectomy was performed by transsphenoidal approach. The operative material was examined by cellular morphology, based on light and electron microscopy and chromosomal ladder formation assay.
The number of apoptotic cells showed features typical for the process as condensation and fragmentation of chromatin, vacuolisation, damage of the inner structure of mitochondria and oligonucleosomal fragmentation of DNA was found in the study material. The results of the studies demonstrated the occurrence of lanreotide induced apoptosis in somatotropinoma type tumours. The grade of apoptotic cells in macroadenomas type adenomas, treated with lanreotide, ranged from 4.0% to 17.1% (mean 8.7+/-2.6%). Patients in whom hypophysectomy was not preceded by lanreotide treatment demonstrated low level of apoptotic cells (no more than 3.5%).
The studies showed that treatment with somatostatine analogue-lanreotide induced apoptosis of tumour cell suggesting that the induction of apoptotic cell death may be involved in the anticancer activity of somatostatine analogue.
Neuro endocrinology letters 11/2003; 24(5):334-8. · 1.30 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Atrial natriuretic factor (ANF) receptor guanylate cyclase (ANF-RGC) is a single chain transmembrane-spanning protein, containing both ANF binding and catalytic activities. ANF binding to the extracellular receptor domain activates the cytosolic catalytic domain, generating the second messenger cyclic GMP. Obligatory in this activation process is an intervening transduction step, which is regulated by the binding of ATP to the cyclase. The partial structural motif of the ATP binding domain of the cyclase has been elucidated and has been termed ATP Regulatory Module (ARM). The crystal structures of the tyrosine kinase domains of the human insulin receptor and haematopoietic cell kinase were used to derive a homology-based model of the ARM domain of ANF-RGC. The model identifies the precise configuration of the ATP-binding pocket in the ARM domain, accurately represents its ATP-dependent features, and shows that the ATP-dependent transduction phenomenon is a two-step mechanism. In the first step, ATP binds to its pocket and changes its configuration; in the second step, via an unknown protein kinase, it phosphorylates the cyclase for its full activation.
Molecular and Cellular Biochemistry 10/2000; 214(1):7-14. · 2.06 Impact Factor
-
-
[show abstract]
[hide abstract]
ABSTRACT: The importance of the second messengers, Ca(2+) and cyclic GMP, for the process of fertilization is well established; the mechanisms for their intracellular regulations in the testes are, however, poorly understood. This study documents the biochemical, molecular, and functional identity of a Ca(2+)-modulated membrane guanylate cyclase transduction machinery in bovine testes. The machinery is both inhibited and stimulated by free Ca(2+) levels. The Ca(2+)-sensor component of the inhibitory mode of the machinery is GCAP1 (guanylate cyclase activating protein type 1) and for the stimulatory mode is S100B. The transduction component is a Ca(2+)-driven rod outer segment membrane guanylate cyclase type 1, ROS-GC1. The cyclase is predominantly expressed in spermatogenic cells. GCAP1 expression is restricted to a small population of spermatogonia, whereas S100B is present in the majority of spermatocytes and spermatids. The expression of GCAP1 and S100B in spermatocytes and spermatids is mutually exclusive.
Journal of Andrology 28(1):50-8. · 2.97 Impact Factor
