ABSTRACT
AIM OF THE STUDY: Large melanoma tumour caused arterial remodelling of the distal part of the great saphenous vein. The metastasis occurred at the site where inguinal lymphadenectomy was previously performed and the proximal part of the great saphenous vein was resected.The aim of this study is the presentation of such a rare observation and literature overview concerning melanoma metastasis and possible stimuli causing remodelling of veins. MATERIAL AND METHODS: Macroscopic and microscopic analyses of the large blood vessel that supplies melanoma were made. The size and structure of the blood vessel was compared with the regular great saphenous vein. RESULTS: The macroscopic examinations allowed us to ascertain that the blood vessel that was identified intraoperatively as the great saphenous vein, has a thick, stiff wall. The microscopic analysis allowed demonstrated that the tunica media was typical for a muscular artery morphology. The morphometric analysis revealed that the blood vessel wall in the area of metastatic tumour was much thicker than the wall of a regular great saphenous vein. CONCLUSIONS: This malignant melanoma skin metastases caused the recanalisation of the great saphenous vein the lumen of which was obliterated during the initial surgical treatment. The metastatic tumour supplied by large blood vessels grew extensively and caused arterial remodelling of the venous wall.
ABSTRACT
The blood count test results of six patients (five male adolescents and one female adult) who were diagnosed with the hemolytic-uremic syndrome are presented. Certain diverse lesions and especially, their different intensity, were observed. They were referred to the clinical process and the time from syndrome occurrence to biopsy.
Subject(s)
Acute Kidney Injury/pathology , Glomerular Basement Membrane/ultrastructure , Hemolytic-Uremic Syndrome/pathology , Acute Kidney Injury/diagnosis , Adult , Biopsy/methods , Child , Female , Hemolytic-Uremic Syndrome/diagnosis , Humans , Infant , Male , Microscopy/methods , Microscopy, Electron/methodsABSTRACT
Special AT-rich sequence-binding protein 1 (SATB1) is a 'genome organizer,' and it has been proposed as a factor that affects the development and progression of various human neoplasms, including colorectal cancer (CRC). This study aimed to compare SATB1 expression in a group of CRC patients and healthy subjects at the mRNA and protein levels. We collected paired tumor tissue and unchanged mucosa of the large intestine from 102 CRC patients as well as 53 biopsies of normal colon mucosa obtained from healthy patients during screening colonoscopy. Tissue samples were quantified for SATB1 mRNA by quantitative PCR, while SATB1 protein expression was determined by Western blotting and immunohistochemistry. SATB1 mRNA level in tumor tissues was over twofolds lower than in samples of corresponding unchanged tissues and fourfolds lower than in biopsies of healthy colon mucosa. Western blotting analysis revealed that SATB1 protein content in tumor and unchanged tissues of CRC patients was over sixfold and fivefolds higher than in biopsies of healthy colon mucosa, respectively. Immunohistochemical staining demonstrated higher nuclear and cytoplasmic SATB1 reactivity in the tumor tissue compared to unchanged mucosa of CRC patients. Despite these differences, SATB1 mRNA, protein, and immunoreactivity levels did not correlate with patients' clinicopathological data and their overall survival, but the latter analysis was limited by a relatively short period of follow-up. In conclusion, we suggest that some as yet unidentified posttranscriptional mechanisms that regulate SATB1 expression may be altered in the CRC tissue.
Subject(s)
Biomarkers, Tumor/biosynthesis , Colorectal Neoplasms/genetics , Matrix Attachment Region Binding Proteins/biosynthesis , RNA, Messenger/biosynthesis , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/genetics , Cell Line, Tumor , Colorectal Neoplasms/pathology , Female , Gene Expression Regulation, Neoplastic , Humans , Kaplan-Meier Estimate , Male , Matrix Attachment Region Binding Proteins/genetics , Middle Aged , Neoplasm Staging , Prognosis , RNA, Messenger/geneticsABSTRACT
The paper presents a summary of immunohistochemical (IHC) and biochemical investigations on the presence of galanin (Gal), one of the neuropeptides abundant in the enteric nervous systems, and three types of its receptors (GalR1-3) in colorectal cancer (CRC) tissue and non-involved colon wall and their associations with clinical-pathological data of the CRC patients. We were the first to morphologically demonstrate the presence of endogenous Gal in CRC sections and measure its content in homogenates of tumor tissue and dissected compartments of unchanged colon wall. The prominent atrophy of myenteric plexuses displaying Gal immunoreactivity (Gal-Ir) located close to the tumor invasion was found to be accompanied by higher Gal content in the tumor-adjacent muscularis externa than in tumor-distant tissue. In further studies for the first time, we demonstrated by the IHC technique the presence of the GalR1-3 receptors in the CRC tumors and the colon mucosa and found that higher GalR3-Ir in the tumor tissue correlated with longer overall survival of CRC patients. Furthermore, we discovered that lower GalR1 expression in submucosal plexuses located near the tumor correlated with a better prognosis in patients with CRC. These findings suggest that GalR1 could be considered as a novel therapeutic target in CRC. In conclusion, our morphological investigations provided novel data documenting the involvement of Gal and its receptors in the progression of CRC and showed the usefulness of the IHC technique for the prognosis of CRC patients.
Subject(s)
Colorectal Neoplasms , Enteric Nervous System , Humans , Galanin , Prognosis , Colorectal Neoplasms/diagnosisABSTRACT
Colorectal cancer is the most common cancer of the gastrointestinal tract. It is considered as a biological model of a certain type of cancerogenesis process in which progression from an early to late stage adenoma and cancer is accompanied by distinct genetic alterations. Clinical and pathological parameters commonly used in clinical practice are often insufficient to determine groups of patients suitable for personalized treatment. Moreover, reliable molecular markers with high prognostic value have not yet been determined. Molecular studies using DNA-based microarrays have identified numerous genes involved in cell proliferation and differentiation during the process of cancerogenesis. Assessment of the genetic profile of colorectal cancer using the microarray technique might be a useful tool in determining the groups of patients with different clinical outcomes who would benefit from additional personalized treatment. The main objective of this study was to present the current state of knowledge on the practical application of gene profiling techniques using microarrays for determining diagnosis, prognosis and response to treatment in colorectal cancer.
Subject(s)
Adenoma/diagnosis , Adenoma/genetics , Colorectal Neoplasms/diagnosis , Colorectal Neoplasms/genetics , Gene Expression Profiling/methods , Genetic Markers/genetics , Oligonucleotide Array Sequence Analysis/methods , Adenoma/therapy , Colorectal Neoplasms/therapy , DNA/genetics , DNA/therapeutic use , Disease Progression , Humans , Medical Oncology/methods , Mutation , Prognosis , Transcriptome , Treatment OutcomeABSTRACT
Inhibitor of nuclear factor kappa B kinase subunit B (IKBKB or IKKß) is a key activator of the nuclear factor κB transcription factor pathway. Increased expression and/or aberrant activity of IKBKB have been observed in various types of human cancer. Three independent techniques, reverse transcriptionquantitative polymerase chain reaction, western blotting and immunohistochemistry, were used to demonstrate that IKBKB expression is decreased in clear cell renal cell carcinoma (ccRCC). Notably, the patients with upregulated IKBKB protein expression were characterized by higher nuclear grade tumors and significantly shorter survival. The findings indicate that IKBKB protein may be of clinical relevance in ccRCC, serving as a marker of poor prognosis and as potential target for adjuvant chemotherapies. Further studies are required to validate the prognostic and predictive value of IKBKB.
Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Renal Cell/pathology , I-kappa B Kinase/metabolism , Kidney Neoplasms/pathology , Adult , Aged , Aged, 80 and over , Carcinoma, Renal Cell/metabolism , Female , Humans , Kidney Neoplasms/metabolism , Male , Middle Aged , Neoplasm Grading , PrognosisABSTRACT
BACKGROUND/AIM: Accumulating evidence suggests that discoidin domain receptor tyrosine kinase 1 (DDR1) has an oncogenic role. Therefore, the aim of this study was to evaluate the potential utility of DDR1 and its post-transcriptional repressors, miR-199a-5p and miR-199b-5p, as prognostic factors in clear cell renal cell carcinoma (ccRCC). PATIENTS AND METHODS: The expression of DDR1 in tumor and normal renal tissues of 56 patients with ccRCC was assessed by reverse transcription quantitative polymerase chain reaction, western blotting and immunohistochemistry. Renal cancer cells were transfected with specific RNA sequences to validate DDR1 as a putative miR-199a/b-5p target. RESULTS: Decreased DDR1 mRNA and protein, as well as miR-199a/b-5p levels were found in ccRCC. Low DDR1 protein was associated with higher nuclear grade and shorter overall survival. DDR1 immunoreactivity was elevated in the nuclei and unchanged in the membrane/cytoplasmic compartment of tumor cells. DDR1 levels correlated with those of miR-199a/b-5p. In addition, we validated DDR1 as a target gene for miR-199a/b-5p in renal cancer cell lines. CONCLUSION: DDR1 expression is altered in ccRCC, but our findings do not support its oncogenic role. In-depth investigation will be necessary to elucidate the exact role and potential utility of miR-199a/b-5p in ccRCC.
Subject(s)
Carcinoma, Renal Cell/secondary , Discoidin Domain Receptor 1/metabolism , Gene Expression Regulation, Neoplastic , Kidney Neoplasms/pathology , MicroRNAs/genetics , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/genetics , Carcinoma, Renal Cell/genetics , Carcinoma, Renal Cell/metabolism , Discoidin Domain Receptor 1/genetics , Female , Follow-Up Studies , Humans , Kidney Neoplasms/genetics , Kidney Neoplasms/metabolism , Male , Middle Aged , Prognosis , Survival Rate , Tumor Cells, CulturedABSTRACT
INTRODUCTION: NKT-like cells are "non-classical", "CD1d-independent" NKT cells which represent highly differentiated, conventional T lymphocytes coexpressing several NK (natural killer) associated receptors. They are effector lymphocytes of both innate and adaptive immunity and simultaneously regulatory cells of the adaptive immune system. They reveal large granular lymphocyte morphology and can mediate both MHC-restricted and MHC-unrestricted cytotoxicity, secrete many cytokines and modulate Th1 immune responses. The aim of our study was to analyze the expression of proteins involved in cellular stress response: sirtuin 1 (SIRT1), heat shock protein 70 (HSP70) and manganese superoxide dismutase (SOD2) in NKT-like cells compared to T lymphocytes during ageing. MATERIAL AND METHODS: The study involved three groups of participants: the oldest seniors (n = 25; aged over 85; mean age 88 ± 0.5 ys), the old (n = 30; aged under 85; mean age 76 ± 0.9 ys) and the young (n = 32; mean age 21 ± 0.3 ys). Whole blood samples were analyzed by flow cytometry to assess the NKT-like (CD3+CD56+) and T (CD3+) cell populations. RESULTS: The group of the oldest seniors differed from the other age groups by much higher percentage of both NKT-like cells and T lymphocytes expressing SIRT1, HSP70 and SOD2. The expression of these proteins correlated positively with the age of the participants. Interestingly, the significantly higher expression of the studied protective proteins; i.e. HSP70 and SOD2 was found in CD3+CD56+ cells compared to CD3+ lymphocytes and this phenomenon concerned all the studied age groups. These differences were not significant regarding the expression of SIRT1; however, the same tendency was noticeable. CONCLUSIONS: The analysis of CD3+ and CD3+CD56+ lymphocytes showed the increase in the number of NKT-like cells and decreased number of T cells in the process of ageing. The increased expression of cellular protective proteins SIRT1, HSP70 and SOD2 in NKT-like and T-lymphocytes of the oldest seniors seems to correspond to longevity and the observed correlations may suggest the involvement of these proteins in establishing cellular homeostasis specific for healthy ageing. Furthermore, the higher expression of the protective proteins in NKT-like cells compared to T lymphocytes may indicate their particular role in the interplay between innate and adaptive immunity responses during the process of ageing.
Subject(s)
Aging/metabolism , Gene Expression Regulation , HSP70 Heat-Shock Proteins/biosynthesis , Natural Killer T-Cells/metabolism , Sirtuin 2/biosynthesis , Superoxide Dismutase/biosynthesis , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Natural Killer T-Cells/cytologyABSTRACT
BACKGROUND/AIM: SATB2 (special AT-rich sequence-binding protein 2) is a DNA-binding protein that is involved in transcriptional regulation and chromatin remodeling. SATB2 protein has been described as a promising novel marker in several human cancers. PATIENTS AND METHODS: This study compared SATB2 expression in tumor and matched unchanged renal tissues collected from 57 patients with clear cell renal cell carcinoma (ccRCC). SATB2 mRNA levels were determined by quantitative polymerase chain reaction, while SATB2 protein expression was estimated by immunohistochemistry. Moreover, the associations between SATB2 expression in ccRCC samples and clinicopathological and survival data of the patients were investigated. RESULTS: The mRNA level of SATB2 was lower in tumor tissues than in samples of corresponding unchanged kidney. Although the average immunoreactivity of SATB2 protein did not differ significantly between cancer cells and epithelial cells of proximal convoluted tubules, the decreased SATB2 expression in tumor specimens inversely correlated with the size of primary tumor and predicted worse patients' outcome. CONCLUSION: The results of the presented study suggest the tumor-suppressing function of SATB2 and that the expression level of this protein can be considered a potential prognostic factor in ccRCC.
Subject(s)
Carcinoma, Renal Cell/metabolism , Kidney Neoplasms/metabolism , Matrix Attachment Region Binding Proteins/biosynthesis , Transcription Factors/biosynthesis , Aged , Carcinoma, Renal Cell/genetics , Carcinoma, Renal Cell/pathology , Case-Control Studies , Cell Growth Processes/physiology , Female , Humans , Immunohistochemistry , Kidney Neoplasms/genetics , Kidney Neoplasms/pathology , Male , Matrix Attachment Region Binding Proteins/genetics , Middle Aged , RNA, Messenger/genetics , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction , Survival Rate , Transcription Factors/geneticsABSTRACT
INTRODUCTION: It have been found previously that colorectal cancer (CRC) is accompanied by atrophy of myenteric plexuses (MPs) localized close to the tumor. The aim of the study was to compare ultrastructure of MPs localized in the unchanged part of the colon wall distant to CRC tumor with the ultrastructure of MPs in the vicinity of CRC tumor. MATERIAL AND METHODS: The present study was conducted using post-operative material derived from 11 patients with CRC. Samples of colon wall were taken from the margin of cancer invasion and from a macroscopically unchanged segment of the large intestine, immediately fixed and processed according to the standard protocol for transmission electron microscopy studies. RESULTS: In the MPs localized in the control part of colon wall the presence of numerous unmyelinated axons and cell bodies of neurons, interstitial cells of Cajal and enteroglial cells were observed. As compared to control samples, in the MPs located close to the tumor invasion, expansion of the extracellular matrix and myelin-like structures accompanying some nerve fibers were found. The appearance of mast and plasma cells was observed within MPs in the vicinity of CRC tumor. Sporadically, apoptotic cells were present inside the MPs. CONCLUSIONS: The presence of myelin-like structures and apoptotic cells within MPs located close to tumor invasion suggests that atrophy of MPs may be caused by factors released from CRC tumor.
Subject(s)
Adenocarcinoma/pathology , Colon/ultrastructure , Colorectal Neoplasms/pathology , Myenteric Plexus/ultrastructure , Adenocarcinoma/ultrastructure , Aged , Colorectal Neoplasms/ultrastructure , Female , Humans , Male , Myenteric Plexus/pathologyABSTRACT
BACKGROUND: Histone acetyltransferase E1A-binding protein p300 (EP300), tumor protein p53 (TP53) and B-cell lymphoma-2-associated X protein (BAX) contribute to the regulation of the cell cycle and apoptosis, cellular processes that are often impaired in cancer cells. The aim of this study was to determine the expression levels of EP300, TP53 and BAX genes and their respective proteins in clear cell renal cell carcinoma (ccRCC) and evaluate the value of these factors as prognostic factors. MATERIALS AND METHODS: EP300, TP53 and BAX expression at the transcript and protein levels were determined by quantitative polymerase-chain reaction (QPCR) and immunohistochemistry (IHC) in paired tumor and kidney specimens from 31 patients with ccRCC. RESULTS: Levels of EP300, TP53 and BAX transcripts were found increased in tumor tissues. Immunoreactivity for TP53 was elevated in cancer cells when compared to unchanged kidney, while EP300 and BAX immunoexpression in ccRCC did not differ from that of normal renal tissue. Immunoreactivity for TP53 was positively associated with larger tumor size. In contrast, stronger BAX immunoexpression correlated with smaller tumor diameters. The average immunoreactivity for BAX was higher in localized, kidney-confined tumor than in advanced/recurrent tumors. None of the analyzed transcripts or proteins correlated with the overall survival of patients. CONCLUSION: Although TP53 and BAX immunoreactivity levels were associated with some clinicopathological parameters of the patients, the expression of EP300, TP53 and BAX did not reveal any prognostic significance in ccRCC.
Subject(s)
Carcinoma, Renal Cell/metabolism , E1A-Associated p300 Protein/metabolism , Kidney Neoplasms/metabolism , Tumor Suppressor Protein p53/metabolism , bcl-2-Associated X Protein/metabolism , Adult , Aged , Aged, 80 and over , Carcinoma, Renal Cell/genetics , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , E1A-Associated p300 Protein/genetics , Female , Humans , Kidney/metabolism , Kidney Neoplasms/genetics , Male , Middle Aged , Prognosis , Transcription Factors/genetics , Transcription Factors/metabolism , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolism , bcl-2-Associated X Protein/geneticsABSTRACT
E1A binding protein P300 (EP300), tumor protein P53 (TP53) and BCL2 associated X, apoptosis regulator (BAX) genes encode proteins which cooperate to regulate important cellular processes. The present study aimed to determine the expression levels of EP300, TP53 and BAX in colorectal cancer (CRC) and to investigate their prognostic value and association with the progression of CRC. Tumor and matched unchanged colorectal tissues were collected from 121 CRC patients. Quantitative polymerase chain reaction and immunohistochemistry were used to assess the mRNA and protein levels of the studied genes. Altered expression of the studied genes in CRC tissues was observed at both the mRNA and protein levels. The depth of invasion was associated with TP53 mRNA levels and was correlated negatively with BAX mRNA expression. Moreover, a relationship between tumor location and BAX mRNA content was noted. BAX immunoreactivity was correlated positively with the intensity of p300 immunostaining and was associated with lymph node involvement and tumor-node-metastasis (TNM) disease stage. Univariate regression analysis revealed that overexpression of p53 and BAX in CRC tissues was associated with poor patient outcome. In conclusion, dysregulation of the expression of the studied genes was found to contribute to CRC pathogenesis. The association between p300 and BAX levels suggests the existence of an interdependent regulatory mechanism of their expression. Moreover, BAX expression may be regulated alternatively, in a p53-independent manner, since the lack of correlations between expression of these factors was observed.
Subject(s)
Biomarkers, Tumor/metabolism , Colorectal Neoplasms/mortality , Colorectal Neoplasms/pathology , E1A-Associated p300 Protein/metabolism , Tumor Suppressor Protein p53/metabolism , bcl-2-Associated X Protein/metabolism , Adult , Aged , Aged, 80 and over , Colorectal Neoplasms/metabolism , Female , Follow-Up Studies , Humans , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Invasiveness , Prognosis , Survival RateABSTRACT
Galanin (GAL) is a 30-amino acid neuropeptide that is expressed in both the central and peripheral nervous system, including the enteric nervous system (ENS). Increased GAL concentrations have been identified in the blood of colorectal cancer (CRC) patients. The aim of the present study was to assess whether sections of the colon wall containing ENS plexuses or CRC tumor are associated with increased GAL concentrations. Blood samples were collected from 68 CRC patients and 39 healthy volunteers. In addition, samples of CRC tumors and colon wall tissue in close proximity to and distant from the neoplastic tissue were obtained from 22 CRC patients. The GAL concentration of sera and tissue homogenates obtained from three sections of the colon wall (mucosa with submucosa, muscularis externa and CRC tumor) was analyzed by ELISA. The localization of GAL was evaluated using immunohistochemistry and morphometry was used to measure the distribution of GAL-immunoreactive (GAL-Ir) myenteric plexuses in the vicinity of cancer invasion and in sections of the colon wall distant from the tumor. The GAL serum concentration of CRC patients was 2.4 times higher than that of the control group. The GAL concentration was highest in the homogenates of neoplastic tissue and mucosa obtained from the control (distant) section of the intestinal wall, followed by that in the mucosa and muscularis externa proximal to the tumor. The lowest GAL concentrations were identified within the muscular layer of the colon wall located distant from the tumor. Strong GAL immunoreactivity was identified in the cancer cells, intestinal epithelium and the submucosal and myenteric plexuses. Morphometric analysis revealed that the GAL-Ir myenteric plexuses in the vicinity of tumor infiltration were significantly smaller in size than those in the intact section of the large intestine. Furthermore, no associations were identified between the clinicopathological characteristics of CRC patients and GAL serum and tissue concentration. The increased GAL serum concentrations observed in CRC patients in comparison to healthy controls may result from GAL secretion by CRC tumors, however, other sources of GAL cannot be excluded. The atrophy of myenteric plexuses within close proximity to the tumor may affect the colon function of CRC patients. In conclusion, investigation into the presence of GAL in the colon wall and serum of CRC patients revealed that serum and tissue GAL levels may present useful potential biomarkers in CRC patients.
ABSTRACT
INTRODUCTION: The previously performed studies showed that the presence of colorectal cancer (CRC) tumor is associated with the atrophy of myenteric plexuses in the vicinity of cancer invasion; however, the possible mechanisms of this phenomenon are not known. The aim of the present study was to determine whether the atrophic changes of the enteric nervous system (ENS) within an intestine wall of the CRC patients were caused by apoptosis or necrosis and whether they were associated with changes in the number of galanin-immunore-active (GAL-Ir) neurons. MATERIAL AND METHODS: Samples of the large intestine wall located close to the CRC invasion and control, distally-located part of the colon, were collected from 9 CRC patients. The size of ENS plexuses and the number of neurons were compared. Triple immunofluorescent staining was used to visualize the co-expression of caspase 3 (CASP3) or caspase 8 (CASP8) with GAL and protein gene-product 9.5 (PGP 9.5, panneuronal marker) in the submucosal and myenteric ENS plexuses. The cells expressing myeloperoxidase (MPO, marker of neutrophils) and CD68 (marker of macrophages) were detected by immunohistochemistry around/in myenteric plexuses (MPs) and in the muscularis externa of the colon wall in the vicinity of tumor invasion. RESULTS: Myenteric plexuses in the vicinity of the CRC tissue were significantly smaller and had lower number of neurons per plexus than distantly located plexuses. The number of CASP8- and CASP3-Ir neurons in the ENS plexuses was similar in the colon wall both close to and distally from tumor invasion. The number of CASP8-Ir neurons within MPs located close to CRC invasion was higher than of CASP3-Ir neurons. The percentage of neurons co-expressing CASP8 and GAL in myenteric plexuses close and distantly from tumor was three-fold lower than of those co-expressing CASP3 and GAL. The mean number of neutrophils and macrophages inside and around myenteric plexuses located close to tumor invasion was higher or similar, respectively, as compared with adjacent muscularis externa. CONCLUSIONS: The atrophy of myenteric plexuses in the vicinity of CRC invasion is not caused by apoptosis or necrosis. The differences in the proportions of neurons expressing galanin and the studied caspases suggest as yet unknown role of this neuropeptide in the mechanisms of neuron's atrophy in MPs located close to CRC tumor.
Subject(s)
Colorectal Neoplasms/pathology , Myenteric Plexus/pathology , Aged , Apoptosis/physiology , Atrophy , Biomarkers, Tumor , Caspase 3/biosynthesis , Caspase 8/biosynthesis , Colorectal Neoplasms/immunology , Colorectal Neoplasms/metabolism , Female , Humans , Immunohistochemistry , Male , Middle Aged , Myenteric Plexus/immunology , Myenteric Plexus/metabolism , Necrosis , Neoplasm Invasiveness , Neurons/metabolism , Neurons/pathology , Peroxidase/biosynthesis , Submucous Plexus/immunology , Submucous Plexus/metabolism , Submucous Plexus/pathology , Ubiquitin Thiolesterase/biosynthesisABSTRACT
BACKGROUND: Pleiomorphic adenoma gene-like 1 (PLAGL1) protein was originally shown to induce cell-cycle arrest and promote apoptosis in several types of human tumors and therefore it was considered a candidate tumor suppressor. The involvement of PLAGL1 gene in the etiology and pathogenesis of clear cell renal cell carcinoma (ccRCC) has not been evaluated. The purpose of the present study was to determine the expression level of PLAGL1 in ccRCC and to investigate its potential utility as a prognostic factor. MATERIALS AND METHODS: We applied quantitative real-time polymerase chain reaction (QPCR), western blotting and immunohistochemistry to measure PLAGL1 mRNA/protein contents in paired tumor and kidney specimens of 40 patients with ccRCC. RESULTS: PLAGL1 mRNA and protein levels were increased in tumor tissues as determined by QPCR and immunohistochemistry, respectively. The average content of PLAGL1 protein measured by western blotting did not differ between tumor and non-cancerous kidney tissues. However, increased PLAGL1 protein level in ccRCC tissue positively correlated with the presence of distant metastases and worse patient outcome. CONCLUSION: These results suggest an oncogenic role of PLAGL1 in the progression of ccRCC and its potential value as a prognostic marker.
Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Renal Cell/secondary , Cell Cycle Proteins/metabolism , Kidney Neoplasms/pathology , Transcription Factors/metabolism , Tumor Suppressor Proteins/metabolism , Biomarkers, Tumor/genetics , Blotting, Western , Carcinoma, Renal Cell/genetics , Carcinoma, Renal Cell/metabolism , Carcinoma, Renal Cell/mortality , Cell Cycle Proteins/genetics , Disease Progression , Female , Follow-Up Studies , Gene Expression Regulation, Neoplastic , Humans , Immunoenzyme Techniques , Kidney Neoplasms/genetics , Kidney Neoplasms/metabolism , Kidney Neoplasms/mortality , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Grading , Neoplasm Staging , Prognosis , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Survival Rate , Transcription Factors/genetics , Tumor Suppressor Proteins/geneticsABSTRACT
BACKGROUND: Altered expression of special AT-rich sequence binding protein 1 (SATB1) was reported in several types of human cancers. This study aimed to determine the expression levels of SATB1, as well as miR-21-5p -the post-transcriptional repressor of SATB1 expression- in clear cell renal cell carcinoma (ccRCC) and to investigate their association with the progression of ccRCC. MATERIALS AND METHODS: Immunohistochemistry and quantitative polymerase chain reaction were used to assess the expression of SATB1 protein and mRNA as well as miR-21-5p in tumor and matched normal kidney tissues collected from 56 ccRCC patients. RESULTS: Nuclear SATB1 immunoreactivity was elevated in ccRCC cells while its cytoplasmic expression was decreased. SATB1 mRNA level was down-regulated in ccRCC tissue and inversely correlated with the content of miR-21-5p. Down-regulation of SATB1 mRNA and up-regulation of miR-21-5p were associated with shorter patient survival. CONCLUSION: Decreased expression of SATB1 in ccRCC may result from over-expressed miR-21-5p. Our data suggest that SATB1 may have a potential value as a prognostic marker in ccRCC.
Subject(s)
Carcinoma, Renal Cell/genetics , Carcinoma, Renal Cell/mortality , Gene Expression Regulation, Neoplastic , Kidney Neoplasms/genetics , Kidney Neoplasms/mortality , Matrix Attachment Region Binding Proteins/genetics , MicroRNAs/genetics , Adult , Aged , Carcinoma, Renal Cell/pathology , Down-Regulation , Female , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Kidney Neoplasms/pathology , Male , Middle Aged , Neoplasm Grading , Neoplasm Metastasis , Neoplasm Staging , Prognosis , Proportional Hazards Models , Protein Transport , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
INTRODUCTION: PLAGL1 (pleiomorphic adenoma gene-like 1) is a C2H2-type zinc finger transcription factor associated with the regulation of cell growth and development. Although PLAGL1 expression in kidney was assessed by biochemical methods, the exact localization of the PLAGL1 protein in human kidney has not yet been described. MATERIAL AND METHODS: Macroscopically unchanged specimens of kidney tissue were collected from 39 patients undergoing nephrectomy due to renal cell carcinoma. H & E staining of paraffin sections was used to assess histology of the kidney whereas immunohistochemistry was used to localize PLAGL1 protein in kidney compartments. In addition, database sequences search for putative PLAGL1 binding sites among the kidney-related genes was performed. RESULTS: PLAGL1 staining intensity differed depending on the kidney compartment. Strong PLAGL1 immunoreactivity was found in thick ascending limbs of Henle's loop, distal tubules and collecting ducts, whereas PLAGL1 expression in proximal tubules and renal corpuscles (including podocytes) was moderate and weak, respectively. By the in sillico screening of promoter sequences for PLAGL1 specific DNA-binding sites GGG-GCCCC we designated 43 candidate genes for PLAGL1-regulated genes. Analysis of their functional annotations identified three significantly over-represented gene sets: inositol phosphate metabolic processes (GO), endocrine and other factor-regulated calcium reabsorption (KEGG) and calcium signaling pathways (KEGG). CONCLUSION: Differences in the renal expression of PLAGL1 suggest that this protein may be involved in the regulation of several cellular pathways both as transcriptional factor and coactivator/corepressor of other tran-scription factors reflecting its role in the cell type-specific control of gene expression.
Subject(s)
Cell Cycle Proteins/metabolism , Kidney Tubules, Collecting/metabolism , Kidney Tubules, Distal/metabolism , Loop of Henle/metabolism , Transcription Factors/metabolism , Tumor Suppressor Proteins/metabolism , Adult , Aged , Aged, 80 and over , Cell Cycle Proteins/genetics , Female , Humans , Male , Middle Aged , Organ Specificity , Transcription Factors/genetics , Tumor Suppressor Proteins/geneticsABSTRACT
Pleomorphic adenoma gene-like 1 gene (PLAGL1) encodes a zinc-finger nuclear transcription factor which promotes apoptosis and cell cycle arrest. Loss or downregulation of its expression has been observed in various human neoplasms. This study compared PLAGL1 expression in colorectal cancer (CRC) tissue and colon mucosa of healthy subjects at the mRNA and protein levels, and estimated its prognostic value. The PLAGL1 mRNA levels were also determined in CRC cell lines. We collected paired tumor tissue and unchanged mucosa of the large intestine from 121 CRC patients as well as 72 colon biopsies of healthy subjects obtained during screening colonoscopy. PLAGL1 mRNA levels were determined by quantitative PCR, while PLAGL1 protein expression was estimated by western blotting and immunohistochemistry. PLAGL1 mRNA level in tumor tissue was ~2-fold lower than in samples of corresponding unchanged tissues and biopsies of healthy colon mucosa. Downregulated expression of PLAGL1 mRNA was also observed in all tested CRC cell lines. Although the average content of PLAGL1 protein did not differ significantly between tumor and unchanged tissues of CRC patients or colon mucosa of healthy individuals, the decreased PLAGL1 protein levels in tumor specimens correlated with lymph node involvement, the presence of metastases and higher TNM disease stage. The PLAGL1 expression level did not correlate significantly with patient overall survival; however, the hazard ratio for patients whose tumor tissues showed reduced PLAGL1 immunohistochemical staining was twice higher than in patients with increased PLAGL1 immunoreactivity. In conclusion, these results suggest that dysregulation of PLAGL1 expression may be involved to some extent in the progression of CRC, but the so far collected patient survival data do not confirm applicability of the PLAGL1 expression level as a prognostic factor in CRC.