cyclin-d1 and Wilms-Tumor

Pediatric renal tumors overlap histomorphologically and may cause misdiagnosis. We aimed to determine the role of immunohistochemical staining of Cyclin D1, PTEN, beta-catenin and PDGFR-alpha on pediatric renal tumors.. Thirty-six cases of 8 different tumors were included in the study. Four blocks of paraffin tissue microarray were constructed. Cyclin D1, PTEN, beta-catenin and PDGFR-alpha were used in all cases. Staining intensity and extent were graded.. All cases of clear cell sarcoma (CCS) and epithelial components of Wilms tumor (WT) showed immunopositivity for Cyclin D1 but blastemal and stromal components of WT were negative. All cases of CCS and most cases of WT consisting of blastemal and stromal components demonstrated loss of expression with PTEN.. Cyclin D1 is not a specific immunohistochemical marker due to its strong and diffuse positivity in CCS cases. It may be useful to differentiate CCS from blastemal and stromal components of WT. Other markers except cyclin D1 do not have a role in the differential diagnosis.

Topics: beta Catenin; Biomarkers, Tumor; Child; Cyclin D1; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Kidney Neoplasms; Male; PTEN Phosphohydrolase; Receptor, Platelet-Derived Growth Factor alpha; Sarcoma, Clear Cell; Wilms Tumor

Clear cell sarcoma of kidney (CCSK) is the second most common pediatric renal malignancy, constituting ∼3% of renal tumors. Due to its morphologic diversity, the diagnosis of CCSK is often challenging. Recent studies have identified internal tandem duplication of BCL6 corepressor (BCOR) gene in CCSKs which coupled with cyclin D1 immunoreactivity, is helpful in differentiating it from its mimics, particularly blastema-rich Wilms tumor (WT), malignant rhabdoid tumor (MRT), and congenital mesoblastic nephroma (CMN). We aimed to evaluate the utility of cyclin D1 and BCOR immunohistochemistry in differentiating CCSK from its morphologic mimics.. Our cohort comprised of 38 pediatric renal tumors which included CCSK (n=18), WT (n=10), MRT (n=5), and CMN (n=5) cases. A detailed clinicopathologic analysis was performed, and tissue microarray were constructed for CCSK and WT, while MRT and CMN tumors were individually stained.. The age ranged from 2 months to 16 years with male:female ratio of 3:1. Strong, diffuse nuclear immunoreactivity for cyclin D1 and BCOR was noted in 61% (n=11/18) and 83% (n=15/18) of CCSK, respectively, while it was significantly less in WT (n=3/10 for cyclin D1) (n=2/10 for BCOR). None of the MRT and CMN examples demonstrated any immunoreactivity. Interestingly, only the blastemal component of WTs showed distinct, rare nuclear immunoreactivity for cyclin D1 or BCOR and the combination of these was never positive in a given case.. Our results provide evidence that concurrent immunopositivity with cyclin D1 and BCOR is helpful in distinguishing CCSK from its morphologic mimics.

Topics: Adolescent; Biomarkers, Tumor; Child; Child, Preschool; Cyclin D1; Diagnosis, Differential; Female; Follow-Up Studies; Humans; Immunohistochemistry; Infant; Kidney Neoplasms; Male; Nephroma, Mesoblastic; Prognosis; Proto-Oncogene Proteins; Repressor Proteins; Rhabdoid Tumor; Sarcoma, Clear Cell; Wilms Tumor

The aim of present report was to elucidate the effect of cell division cycle associated 4 (CDCA4) on the proliferation and apoptosis of Wilm's tumor cells, and to further evaluate its underlying mechanism.. The expression profiles of CDCA4 and clinical information of Wilm's tumor patients were obtained from public Therapeutically Applicable Research to Generate Effective Treatments (TARGET) database portal. Real-time qPCR and western blot analyses were utilized to determine the expression levels of CDCA4. Gain- and loss-of-function of CDCA4 assays were conducted with transfection technology to investigate the biological role of CDCA4 in Wilm's tumor cells. Cell counting kit 8 and flow cytometer assays were employed to examine the effect of CDCA4 on the cells proliferation and apoptosis. Protein expression levels of indicated markers in each group of Wilm's tumor cells were measured by western blot.. The transcriptional expression of CDCA4 was drastically upregulated in Wilm's tumor tissues according to the public TARGET database and in Wilm's tumor cells. The cells viability was remarkably reduced whereas the cells apoptosis was increased in CDCA4-knockdown group compared with negative control group. However, CDCA4-overexpression group promoted the cells proliferation and suppressed the cells apoptosis. Furthermore, the protein expression levels of p-AKT, p-mTOR, and Cyclin D1 were significantly reduced after depletion of CDCA4, whereas overexpression of CDCA4 dramatically elevated these markers' expression levels.. CDCA4 is highly expressed in Wilm's tumor and promoted the proliferation whereas inhibited the apoptosis of Wilm's tumor cells through activating the AKT/mTOR signaling pathway.

Topics: Apoptosis; Cell Cycle Proteins; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cyclin D1; Humans; Kidney Neoplasms; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases; Wilms Tumor

Wilms'tumor is the most common malignant tumor with a poor clinical prognosis because of metastasis or recurrence among children worldwide. CD151, a member of transmembrane 4 superfamily, has now been confirmed to be involved in tumor progression including the proliferation, migration, invasion and metastasis of tumor cells. GSK-3β/P21/cyclinD signaling pathway plays a critical role in the cell cycle progression, regulating cellular proliferation. In this study, CD151 protein and mRNA levels were examined by western blot and RT-PCR. The proliferation of SK-NEP-1 cells was examined by CCK8 assay and the migration of SK-NEP-1 cells was detected with wound healing assay. Furthermore, p-GSK3β protein, GSK3β protein, p21protein and CyclinD protein were examined by western blot to verify whether CD151 could regulate the Wilms'tumor progression via the GSK-3β/P21/cyclinD signaling pathway. The RT-PCR and western blot results showed that CD151 protein was upregulated in Wilms'tumor cells compared with the control. The results by CCK8 assay and wound healing assay demonstrated that CD151 overexpression promoted the proliferation and migration in SK-NEP-1 cells and CD151 interference showed the opposite effects. Western blot assay revealed that CD151 activated the GSK-3β/P21/cyclinD signaling pathway and upregulated the expression of p-GSK3β protein, p21protein and CyclinD protein. It was also verified that CD151 promotes proliferation and migration of SK-NEP-1 cells through the GSK-3β/P21/cyclinD signaling pathway in this study. The specific aim of the study is to investigate and verify the role of CD151 in Wilm's tumor. Therefore, in-depth study on the molecular mechanisms will provide new strategies and methods for the treatment of Wilm's tumor.

Topics: Cell Line, Tumor; Cell Movement; Cell Proliferation; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Glycogen Synthase Kinase 3 beta; Humans; Kidney Neoplasms; Signal Transduction; Tetraspanin 24; Wilms Tumor

Small round blue cell tumors (SRBCTs) of children and adolescents are often diagnostically challenging lesions. With the increasing diagnostic approach based on small biopsies, there is the need of specific immunomarkers that can help in the differential diagnosis among the different tumor histotypes to assure the patient a correct diagnosis for proper treatment. Based on our recent studies showing cyclin D1 overexpression in both Ewing sarcoma/primitive peripheral neuroectodermal tumor (EWS/pPNET) and peripheral neuroblastic tumors (neuroblastoma and ganglioneuroblastoma), we immunohistochemically assessed cyclin D1 immunoreactivity in 128 cases of SRBCTs in children and adolescents to establish its potential utility in the differential diagnosis. All cases of EWS/pPNET and the undifferentiated/poorly differentiated neuroblastomatous component of all peripheral neuroblastic tumors exhibited strong and diffuse nuclear staining (>50% of neoplastic cells) for cyclin D1. In contrast, this marker was absent from rhabdomyosarcoma (regardless of subtype) and lymphoblastic lymphoma (either B- or T-cell precursors), whereas it was only focally detected (<5% of neoplastic cells) in some cases of Wilms tumor (blastemal component) and desmoplastic small round cell tumor. Our findings suggest that cyclin D1 can be exploitable as a diagnostic adjunct to conventional markers in confirming the diagnosis of EWS/pPNET or neuroblastoma/ganglioneuroblastoma. Its use in routine practice may also be helpful for those cases of SRBCT with undifferentiated morphology that are difficult to diagnose after application of the conventional markers.

Topics: Adolescent; Biomarkers, Tumor; Biopsy; Bone Neoplasms; Cell Differentiation; Child; Child, Preschool; Cyclin D1; Desmoplastic Small Round Cell Tumor; Diagnosis, Differential; Female; Ganglioneuroblastoma; Humans; Immunohistochemistry; Infant; Kidney Neoplasms; Male; Neuroblastoma; Neuroectodermal Tumors, Primitive, Peripheral; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Predictive Value of Tests; Retrospective Studies; Rhabdomyosarcoma; Sarcoma, Ewing; Wilms Tumor; Young Adult

Distinguishing clear cell sarcoma of the kidney (CCSK) from other paediatric malignancies, particularly blastema-rich Wilms tumour (WT) and congenital mesoblastic nephroma (CMN), is challenging. Specific immunohistochemistry for CCSK does not exist, and diagnosis rests upon histopa thology. Recently, the YWHAE-FAM22 rearrange ment, identical to that in endometrial stromal sarcoma (ESS), has been identified in CCSKs. As this fusion results in overexpression of cyclin D1 in ESS, we postulated that overexpression would also occur in CCSK; cyclin D1 immunohistochemistry could then be used to differentiate CCSK from other tumours. The goal of this study was therefore to evaluate the utility of cyclin D1 immunohistochemistry in identifying CCSK and helping to differentiate it from its mimics.. Cyclin D1 expression was evaluated in 59 renal tumours-CCSK (14), WT (25), rhabdoid tumour (four), Ewing sarcoma (five), and CMN (11)-and four neuroblastomas. All 14 CCSKs showed diffuse and strong reactivity. In contrast, the blastematous component of most WTs showed only rare positive nuclei, that of rhabdoid tumours showed rare to focal immunoreactivity, and that of more than half of CMNs showed weak or focal immunoreactivity. Most Ewing sarcomas and all neuroblastomas showed diffuse moderate to strong staining.. Cyclin D1 is most helpful in distinguishing CCSK from WT, rhabdoid tumour, and some CMNs, but not from neuroblastoma or Ewing sarcomas.

Topics: Biomarkers, Tumor; Child, Preschool; Cyclin D1; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Kidney Neoplasms; Male; Nephroma, Mesoblastic; Neuroblastoma; Rhabdoid Tumor; Sarcoma, Clear Cell; Sarcoma, Ewing; Wilms Tumor

Loss of heterozygosity (LOH) in 16q appears in ~20-30% cases of Wilms' tumor. Within this region, known as common fragile site FRA16D, the WWOX tumor suppressor gene is located. Abnormalities of WWOX gene expression levels were observed in many tumor types and were associated with worse prognosis. The purpose of this study was to investigate the role of the WWOX tumor suppressor gene in Wilms' tumor samples. We evaluated the correlation between expression of WWOX and genes involved in proliferation (Ki67), apoptosis (BCL2, BAX), signal transduction (ERBB4, ERBB2, EGFR), cell cycle (CCNE1, CCND1), cell adhesion (CDH1) and transcription (TP73) using real-time RT-PCR in 23 tumor samples. We also analyzed the potential causes of WWOX gene expression reduction i.e., promoter methylation status (MethylScreen method) and loss of heterozygosity (LOH) status. We revealed a positive correlation between WWOX expression and BCL2, BCL2/BAX ratio, EGFR, ERBB4 isoform JM-a, TP73 and negative correlation with both cyclins. Loss of heterozygosity of the WWOX gene was observed only at intron 8, however, it had no influence on the reduction of its expression levels. Contrary to LOH, methylation of the region covering the 3' end of the promoter and part of exon 1 was associated with statistically significant reduction of WWOX gene expression levels. In the present study we reveal that in Wilms' tumors the WWOX expression levels are positively associated with the process of apoptosis, signal transduction through the ErbB4 pathway and EGFR and negatively with the regulation of the cell cycle (by cyclin E1 and D1). Moreover, our analysis indicates that in this type of tumor the expression of the WWOX gene can be regulated by an epigenetic mechanism--its promoter methylation.

Topics: Apoptosis; bcl-2-Associated X Protein; Cell Proliferation; Child; Child, Preschool; Cyclin D1; Cyclin E; DNA Methylation; ErbB Receptors; Exons; Gene Expression Regulation, Neoplastic; Humans; Infant; Ki-67 Antigen; Kidney Neoplasms; Loss of Heterozygosity; Oncogene Proteins; Oxidoreductases; Promoter Regions, Genetic; Receptor, ErbB-4; Signal Transduction; Tumor Suppressor Proteins; Wilms Tumor; WW Domain-Containing Oxidoreductase

The expression status of the three cyclin D genes (CCND1, CCND2 and CCND3), the two cyclin D-dependent kinase genes (CDK4 and CDK6) and the p16(INK4a) gene was studied in a series of 47 Wilms' tumors, 16 normal mature kidneys and two fetal kidneys. We showed predominant overexpression of CCND2 and CDK4 compared to CCND1/D3 and CDK6 respectively. We found a specific correlation between relapse and CDK4 overexpression, but not CDK6 overexpression. We did not identify any methylation of the p16(INK4a) promoter. This suggests that dysregulation of CCND2 and CDK4 plays a specific role in WT tumorigenesis.

Topics: Cyclin D1; Cyclin D2; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase 6; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinases; Cyclins; DNA Methylation; Humans; Kidney; Kidney Neoplasms; Promoter Regions, Genetic; Proto-Oncogene Proteins; Wilms Tumor