Gene expression profiling of giant cell tumor of bone reveals downregulation of extracellular matrix components decorin and lumican associated with lung metastasis (original) (raw)
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The American Journal of Pathology, 2011
Giant cell tumor of bone can be locally aggressive and occasionally can metastasize in the lungs. To identify new markers predictive of aggressive behavior, we analyzed five patients who developed lung metastasis and five who remained disease free for a minimum of 5 years. Using two-dimensional electrophoresis, we detected 28 differentially expressed spots. Fourteen spots were identified using mass spectrometry, including seven up-regulated and seven down-regulated in metastatic samples and classified according to functional categories. We then selected five proteins involved in cell cycle or apoptosis. Thioredoxin peroxidase, allograft inflammatory factor 1, and ubiquitin E2N had more than threefold up-regulation; glutathione peroxidase 1 had 1.9-fold up-regulation; and heat shock protein 27 showed down-regulation in metastatic samples with a very low P value. After validation and analysis of protein levels, evaluation of clinical impact was assessed in a much wider cohort of primary archival specimens. Immunodetection showed a higher frequency of thioredoxin peroxidase, allograft inflammatory factor 1, ubiquitin E2N, and glutathione peroxidase 1 overexpression in primary tumors that developed into lung metastases or that locally relapsed than in the disease-free group, with variable stain intensity and distribution. Kaplan-Meier analysis showed that high expression of glutathi-Supported by grants from the Italian Ministry of Health (Programma straordinario Ricerca Oncologica) and Istituto Superiore della Sanità (Italia-USA Project and ACC10). Istituto Ortopedico Rizzoli, Centro de Investigación del Cancer-IBMCC and Semmelweis University are partners of the Euro-BoNeT consortium, a network of excellence granted by the European Commission for studying the pathology and genetics of bone tumors.
Journal of Cancer, 2015
Though benign, giant cell tumor of bone (GCTB) can become aggressive and can exhibit a high mitotic rate, necrosis and rarely vascular invasion and metastasis. GCTB has unique histologic characteristics, a high rate of multinucleated cells, a variable and unpredictable growth potential and uncertain biological behavior. In this study, we sought to identify genes differentially expressed in GCTB, thus building a molecular profile of this tumor. We performed quantitative real-time polymerase chain reaction (qPCR), immunohistochemistry and analyses of methylation to identify genes that are putatively associated with GCTB. The expression of the ADAM23 and CDKN2A genes was decreased in GCTB samples compared to normal bone tissue, measured by qPCR. Additionally, a high hypermethylation frequency of the promoter regions of ADAM23 and CDKN2A in GCTB was observed. The expression of the MAP2K3, MMP14, TIMP2 and VIM genes was significantly higher in GCTB than in normal bone tissue, a fact that was confirmed by qPCR and immunohistochemistry. The set of genes identified here furthers our understanding of the molecular basis of GCTB.
Journal of Orthopaedic Research, 2015
Giant cell tumor of bone (GCTb) represents 5% of bone tumors, and although considered benign, 5% metastasize to the lung. The expression of proteins directly or indirectly associated with osteolysis and tumor growth was studied on 163 samples of GCTb. Of these, 33 patients developed lung metastasis during follow-up. The impact of tumor-host interaction on clinical aspects was evaluated with the aim of finding specific markers for new biological therapies, thus improving clinical management of GCTb. Protein expression was evaluated by immunohistochemical analysis on Tissue Microarray. The majority of GCTb samples from patients with metastatic disease were strongly positive to RANKL and its receptor RANK as well as to CAII and MMP-2 and to pro-survival proteins NFIB and c-Fos. Kaplan-Meier analysis indicated a significant difference in metastasis free survival curves based on protein staining. Interestingly, the statistical correlation established a strong association between all variables studied with a higher t coefficient for RANK/RANKL, RANK/NFIB, and RANKL/NFIB pairs. At multivariate analysis co-overexpression of NFIB, RANK and RANKL significantly increased the risk of metastasis with an odds ratio of 13.59 (95%CI 4.12-44.82; p < 0.0005). In conclusion, the interconnection between matrix remodeling and tumor cell activity may identify tumor-host endpoints for new biological treatments.
Proteases and interleukin-6 gene analysis in 92 giant cell tumors of bone
Annals of Oncology
Giant cell tumor of bone (GCT) is a benign tumor with a significant tendency to recur locally and rarely to produce pulmonary metastases. It is characterized by the presence of multinucleated osteoclast-like giant cells together with mononuclear spindle-shaped cells. Few prognostic markers have been reported to predict the clinical outcome of GCT patients, so is very important to find the factor that can be implicated in its potential aggressiveness. Different groups of GCT patients were selected for this study, including patients without evidence of disease and patients who recurred locally or with lung metastasis. The total of 92 tumor samples also included the specimens of the local recurrences and the lung metastases. By using immunohistochemistry and real-time quantitative polymerase chain reaction techniques, the genetic and proteic analyses were performed on the urokinase-type plasminogen activator (u-PA), its receptor (u-PAR) and its inhibitor (PAI-1), which have been descri...
MicroRNA expression profiles in metastatic and non-metastatic giant cell tumor of bone
Histology and histopathology, 2013
Giant cell tumor of bone (GCTB) is a skeletal neoplasm, a locally aggressive tumor that occasionally metastasizes to the lungs. To identify novel biomarkers associated with GCTB progression and metastasis, we performed a miRNA microarray on ten primary tumors of GCTB, of which five developed lung metastases and the rest remained metastasis-free. Between metastatic and non-metastatic GCTB, 12 miRNAs were differentially expressed (such as miR-136, miR-513a-5p, miR-494, miR-224, and miR-542-5p). A decreased level of miR-136 in metastatic versus non-metastatic GCTB was significantly confirmed by the quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) (p=0.04). To identify potential target genes for the differentially expressed miRNAs, we used three target prediction databases. Then, to functionally validate the potential target genes of the differentially expressed miRNAs, we re-analyzed our previous gene expression data from the same ten patients. Eight genes such as...
Gene expression in giant-celltumors
Journal of Laboratory and Clinical Medicine, 2004
Malignant transformation is thought to be associated with changes in the expression of a number of genes, and this alteration in gene expression is considered critical to the development of the malignant phenotype. In this study, gene expression in 8 samples of giant-cell tumor (GCT) of bone, as well as in bone at the site of osteoarthritis and in a variety of normal tissues, was determined at Gene Logic Inc (Gaithersburg, Md) with the use of Affymetrix GeneChip ® U_133 arrays containing approximately 40,000 genes/expressed sequence tags (ESTs). Gene-expression analysis was performed with the use of the Gene Logic GeneExpress ® Software System. Differences in gene expression between GCTs and bone were observed. In addition, genes expressed uniquely in GCTs among these and 519 samples from 20 other tissue types were identified. Some of the genes that were found to be overexpressed in GCTs, such as tartrate-resistant acid phosphatase and the lysosomal H ؉ -transporting ATPase, are also expressed by osteoclasts. Osteoprotegrin ligand (OPGL) was also selectively overexpressed in GCTs. The genes found to be overexpressed in GCTs appear to reflect the genetic profile of osteoclast-lineage cells and also the genetic profile of an osteoclastogenic environment. The genes identified in this study may play a role in the pathogenesis of GCTs, confirm the likely importance of OPGL in GCT pathogenesis, and may indicate other possible targets to which antitumor therapy could be directed. (J Lab Clin Med 2004;144:193-200) Abbreviations: CCR1 ϭ chemokine (C-C motif) receptor 1; cDNA ϭ complementary DNA; cRNA ϭ complementary RNA; CSF ϭ colony-stimulating factor; CSF1R ϭ CSF-1 receptor; DC ϭ dendritic cell; DCIR ϭ dendritic-cell immunoreceptor; EST ϭ expressed sequence tag; EXT1 ϭ multiple extoses 1; EXT2 ϭ multiple extoses 2; GCT ϭ giant-cell tumor; HGK ϭ mitogenactivated protein kinase-4; KCNN4 ϭ potassium intermediate/small conductance calciumactivated channel, subfamily N, member 4; MATK ϭ megakaryocyte-associated tyrosine kinase; MFH ϭ malignant fibrous histiocytomas; MMP ϭ matrix metalloproteinase; NFAT ϭ nuclear factor of activated T-cells; NFATC1 ϭ nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 1; OPGL ϭ osteoprotegrin ligand; RANKL ϭ receptor activator of NF-B ligand; SLC4A2 ϭ solute-carrier family 4, anion-exchanger member 2; TGF ϭ transforming growth factor; TM7SF1 ϭ transmembrane 7 superfamily 1; TRAP ϭ tartrate-resistant acid phosphatase G iant-cell tumor of bone, also known as osteoclastoma, is a primary osteolytic bone neoplasm in which monocytic macrophage/osteoclast precursor cells and multinucleated osteoclastlike giant cells infiltrate the tumor. 1-3 Although benign, GCTs are locally aggressive and most commonly occurs in the epiphyses of long bones. Walker first dem-onstrated that osteoclasts are derived from monocytic osteoclast precursors in blood 4 -8 and provided the rationale for the treatment of human Albers-Schoenberg marble-bone disease with bone-marrow transplantation. The origin of GCT is unknown, but the tumor cells of GCT have been reported to produce chemoattractants that can attract osteoclasts and osteoclast precursors. 3
Gene expression profiling identifies p63 as a diagnostic marker for giant cell tumor of the bone
Modern Pathology, 2008
Giant cell tumor of the bone (GCTOB) is a primary bone tumor that occurs mainly in young adults and is capable of locally aggressive growth. Its histologic appearance can resemble a number of benign and malignant tumors but no useful diagnostic marker is known currently. To identify diagnostic markers for this tumor, global gene expression profiling using cDNA microarray was performed on 6 fresh-frozen GCTOB, 3 aneurysmal bone cysts, 4 fibrous dysplasias and 12 giant cell tumors of tendon sheath/diffuse-type giant cell tumors. Unsupervised hierarchical clustering separated the tumors based on their histopathologic types, and significance analysis of microarray identified several genes including TP73L (encoding the p63 protein) that are significantly highly expressed in GCTOB relative to these other tumors. The diagnostic utility of p63 was subsequently confirmed using anti-p63 antibody on a series of 26 GCTOB, 25 aneurysmal bone cysts, 15 chondroblastomas, 13 giant cell reparative granulomas, 13 chondromyxoid fibromas, 4 brown tumors, 4 fibrous dysplasias, 53 giant cell tumors of tendon sheath/diffuse-type giant cell tumors and 385 additional mesenchymal tumors in tissue microarrays. Strong p63 nuclear staining was present in 18 of 26 (69%) GCTOB, 3 of 15 (20%) chondroblastomas and in 1 of 25 (4%) aneurysmal bone cysts while none of the other tumors commonly considered in the differential diagnosis of GCTOB showed any detectable p63 staining. Strong p63 staining is rare in bone and soft-tissue tumors in general. In contrast to the pattern of p63 staining, the majority of the chondroblastomas (70%) demonstrated S-100 immunoreactivity while only a minority of the GCTOB (8%) was immunoreactive for S-100. These findings altogether show that p63 can be used as a diagnostic marker to aid the clinical diagnosis of GCTOB.
p53 Expression Immuno-histochemistry Index in Stage III Giant Cell Tumor of the Bone
JMED Research, 2014
We analyzed the p53 expression of 50 consecutive cases of stage III giant cell tumor to determine the clinico-pathological correlation. There were 29 male patients compared to 21 females. The mean age was 36.42 years and ranged from 19 to 66 years. Tumors were located mainly around the knee (48%) followed by distal radius (28%). Sixteen percent (16%) had local recurrence, and twenty four percent (24%) presented with or eventually developed lung metastasis. There were 3 patients who had both local recurrence and pulmonary metastasis. The expression of p53 antigen was evaluated by immuno-histochemical staining techniques, using the avidin-biotin perioxidase complex method, using an LSAB2 kit (Dako, Glostrup, Denmark). The scoring was done by counting the positively stained cell in the background of 1000 mononuclear stromal cells. Data was analyzed using PASW version 18.0. The mean value of p53 index obtained as a percentage of 1000 background cells was 37.68 (ranged 1.00-20.0), with standard deviation of 28.33. The p53 index of recurrent tumor was 65.38 compared to 32.42 of those without recurrence, and was statistically significant (mean difference of 32.97 with p value in independent t test of 0.002). The p53 index was also statistically significant in the presence of pulmonary metastases with the mean value of 66.92 compared to 28.45 of those without metastasis (mean difference of 38.47 with p value in independent t test of 0.001). In conclusion, p53 expression is a good prognostic marker to predict the risk of local recurrence and lung metastasis in the giant cell tumor of the bone.
Giant cell tumour of bone (GCTB) is considered to be a histologically benign osseous tumour, which tends to exhibit local invasion and tendency for formation of pulmonary metastases. It has a high rate of recurrence and may undergo malignant transformation. The tumour is composed of three types of cells: osteoclast-like multinucleated giant cells, spindle-shaped stromal cells of mesenchymal origin and cells of the monocyte-macrophage lineage. Giant and stromal cells are responsible for the production and release of matrix met-alloproteinases (MMP-s), which accelerate the breakdown of the extracellular matrix (ECM) and have a key role in local tumour invasion, as well as distant metastasis. The biological behaviour of GCTB can be assessed by radio-graphic appearance on the basis of Campanacci's classification, which classifies the tumour into 3 grades depending on the radiographic signs of local invasion. The aim of the present study was to establish whether a correlation exists between the radiographic appearance of GCTB grades 1 and 3 according to Campanacci's classification and the expression of MMP-9. We studied clinical material from 8 patients and conducted an immunohistochemical analysis of the MMP-9 expression. A positive reaction was observed predominantly in stromal cells and was more intensive in lesions with aggressive radiographic appearance as compared to those with latent behaviour. The established correlation between MMP-9 expression by tumour cells and biological behaviour, assessed on the basis of radiographic appearance, may further support the role 1757