Heparanase isoform expression and extracellular matrix remodeling in intervertebral disc degenerative disease

OBJECTIVE: To determine the molecules involved in extracellular matrix remodeling and to identify and quantify heparanase isoforms present in herniated and degenerative discs. INTRODUCTION: Heparanase is an endo-beta-glucuronidase that specifically acts upon the heparan sulfate chains of proteoglycans. However, heparanase expression in degenerative intervertebral discs has not yet been evaluated. Notably, previous studies demonstrated a correlation between changes in the heparan sulfate proteoglycan pattern and the degenerative process associated with intervertebral discs. METHODS: Twenty-nine samples of intervertebral degenerative discs, 23 samples of herniated discs and 12 samples of non-degenerative discs were analyzed. The expression of both heparanase isoforms (heparanase-1 and heparanase-2) was evaluated using immunohistochemistry and real-time RT-PCR analysis. RESULTS: Heparanase-1 and heparanase-2 expression levels were significantly higher in the herniated and degenerative discs in comparison to the control tissues, suggesting a possible role of these proteins in the intervertebral degenerative process. CONCLUSION: The overexpression of heparanase isoforms in the degenerative intervertebral discs and the herniated discs suggests a potential role of both proteins in the mediation of inflammatory processes and in extracellular matrix remodeling. The heparanase-2 isoform may be involved in normal metabolic processes, as evidenced by its higher expression in the control intervertebral discs relative to the expression of heparanase-1

Análise comparativa histopatológica entre a hérnia de disco contida e extrusa Análisis comparativo histopatológico entre hernia discal contenida y extruida Comparative histopathologic analysis of contained and extruded disc herniation

OBJETIVO: Nosso estudo tem o objetivo de estudar as alterações histopatológicas, tais como neovascularização, infiltrado inflamatório, celularidade, apoptose, degeneração mucoide, alterações granulares e calcificação presentes nos tipos de hérnia (contida e extrusa), e também avaliar essas diferenças entre o núcleo pulposo e ânulo fibroso. MÉTODOS: Foram analisados 65 discos lombares, os quais foram divididos em três grupos: hérnia extrusa com 25 casos, hérnia contida com 28 casos e 12 discos sem alteração degenerativa. Os fragmentos removidos foram separados em ânulo fibroso e núcleo pulposo. Foi realizada análise semiquantitativa por microscopia óptica das alterações histopatológicas. RESULTADO: Em relação aos parâmetros avaliados na análise comparativa entre ânulo fibroso e núcleo pulposo não houve variação estatística significativa entre os grupos, o que mostra que ambas as regiões são semelhantes. A hérnia extrusa apresentou maior proporção de infiltrado inflamatório e neovascularização. As alterações degenerativas não apresentaram uma variação significante conforme o tipo de hérnia. CONCLUSÕES: Na hérnia de disco há uma relação entre neovascularização, infiltrado inflamatório e o tipo de hérnia. Não há diferença histopatológica em relação à porção do disco intervertebral analisada.OBJETIVO: Nuestro estudio tiene el objetivo de analizar las alteraciones histopatológicas tales como neovascularización, infiltrado inflamatorio, celularidad, apoptosis, degeneración mucoide, alteraciones granulares y calcificación, presentes según los tipos de hernia (contenida y extruida), así como evaluar esas diferencias entre el núcleo pulposo y el anillo fibroso. MÉTODOS: Se analizaron 65 discos lumbares, que se dividieron en tres grupos: hernia extruida en 25 casos, hernia contenida en 28 casos y 12 discos sin alteración degenerativa. Los fragmentos extraídos se separaron en anillo fibroso y núcleo pulposo. Se llevó a cabo un análisis semicuantitativo por microscopia óptica de las alteraciones histopatológicas. RESULTADO: Respecto a los parámetros evaluados en el análisis comparativo entre anillo fibroso y núcleo pulposo, no se produjo variación estadística significativa entre los grupos, lo cual muestra que ambas regiones son semejantes. La hernia extruida presentó una mayor proporción de infiltrado inflamatorio y neovascularización. Las alteraciones degenerativas no presentaron una variación significativa según el tipo de hernia. CONCLUSIONES: En la hernia discal hay una relación entre neovascularización, infiltrado inflamatorio y tipo de hernia. No hay diferencia histopatológica respecto a la parte del disco intervertebral analizada.OBJECTIVE: to investigate histopathological changes such as neovascularization, inflammatory infiltrate, cellularity, apoptosis, mucoid degeneration, granular changes, and calcification in contained and extruded disc herniations, and to compare these differences in the nucleus pulposus and annulus fibrosus. METHODS: 65 lumbar discs were evaluated. These were divided in three groups: 25 cases of extruded herniated discs, 28 cases of contained herniated discs, and 12 cases of discs without degenerative changes. Fragments were removed and separated into annulus fibrosus and nucleus pulposus. Semi-quantitative analysis of histopathologic changes was carried out, using a microscope. RESULTS: in the comparative analysis between annulus fibrosus and nucleus pulposus, no statistical differences were obtained between these groups, showing that both regions are similar. The extruded disc herniation presented a higher proportion of inflammatory infiltrate and neovascularization. Degenerative changes did not present significant variation in relation to disc herniation type. CONCLUSION: There is a relation in disc herniation between neovascularitazion, inflammatory infiltrate and type of disc herniation. There is no histopathologic difference in relation of the portion of intervertebral disc analyzed

Crosstalk between tumor cells and lymphocytes modulates heparanase expression

Abstract Background Heparanase (HPSE) is an endo-beta-glucuronidase that degrades heparan sulfate (HS) chains on proteoglycans. The oligosaccharides generated by HPSE promote angiogenesis, tumor growth and metastasis. Heparanase-2 (HPSE2), a close homolog of HPSE, does not exhibit catalytic activity. Previous studies have demonstrated that serum or plasma from breast cancer patients showed increased expression of both heparanases in circulating lymphocytes. The aim of this study was to better understand the mechanisms involved in the upregulation of heparanases in circulating lymphocytes. Methods Lymphocytes collected from healthy women were incubated in the presence of MCF-7 breast cancer cells (co-culture) to stimulate HPSE and HPSE2 overexpression. The protein level of heparanases was evaluated by immunocytochemistry, while mRNA expression was determined by quantitative RT-PCR. Results The medium obtained from co-culture of MCF-7 cells and circulating lymphocytes stimulated the expression of HPSE and HPSE2. Previous treatment of the co-culture medium with an anti-heparan sulfate proteoglycan antibody or heparitinase II inhibited the upregulation of heparanases in circulating lymphocytes. The addition of exogenous heparan sulfate (HS) enhanced the expression of both heparanases. Moreover, the co-cultured cells, as well as MCF-7 cells, secreted a higher number of exosomes expressing an increased level of HS compared to that of the exosomes secreted by circulating lymphocytes from women who were not affected by cancer. Conclusions The results revealed that HS is likely responsible for mediating the expression of heparanases in circulating lymphocytes. HS secreted by tumor cells might be carried by exosome particles, confirming the key role of tumor cells, as well as secreted HS, in upregulating the expression of heparanases, suggesting a possible mechanism of crosstalk between tumor cells and circulating lymphocytes

Expression of heparanase in basal cell carcinoma and squamous cell carcinoma

Abstract: Background: Heparanase is an enzyme that cleaves heparan sulfate chains. Oligosaccharides generated by heparanase induce tumor progression. Basal cell carcinoma and squamous cell carcinoma comprise types of nonmelanoma skin cancer. Objectives: Evaluate the glycosaminoglycans profile and expression of heparanase in two human cell lines established in culture, immortalized skin keratinocyte (HaCaT) and squamous cell carcinoma (A431) and also investigate the expression of heparanase in basal cell carcinoma, squamous cell carcinoma and eyelid skin of individuals not affected by the disease (control). Methods: Glycosaminoglycans were quantified by electrophoresis and indirect ELISA method. The heparanase expression was analyzed by quantitative RT-PCR (qRTPCR). Results: The A431 strain showed significant increase in the sulfated glycosaminoglycans, increased heparanase expression and decreased hyaluronic acid, comparing to the HaCaT lineage. The mRNA expression of heparanase was significantly higher in Basal cell carcinoma and squamous cell carcinoma compared with control skin samples. It was also observed increased heparanase expression in squamous cell carcinoma compared to the Basal cell carcinoma. Conclusion: The glycosaminoglycans profile, as well as heparanase expression are different between HaCaT and A431 cell lines. The increased expression of heparanase in Basal cell carcinoma and squamous cell carcinoma suggests that this enzyme could be a marker for the diagnosis of such types of non-melanoma cancers, and may be useful as a target molecule for future alternative treatment

Heparanase Expression in Circulating Lymphocytes of Breast Cancer Patients Depends on the Presence of the Primary Tumor and/or Systemic Metastasis1

Heparanase is an endo-β-glucuronidase that is capable of degrading heparan sulfate chains of proteoglycans, generating a variety of bioactive molecules such as growth factors and chemotactic and angiogenic agents. The expression of heparanase was investigated in the peripheral blood mononuclear cell fraction (PBMC) of 30 patients with breast cancer and 20 healthy control women by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. PBMC samples from all breast cancer patients at study entry showed the expression of heparanase, whereas no expression was observed for healthy women. Immunocytochemistry analysis demonstrated that heparanase was expressed in lymphocytes of breast cancer PBMC. Throughout follow-up, heparanase expression by RTPCR decreased significantly after surgery in patients treated with neoadjuvant chemotherapy (P = .002) and after tamoxifen treatment (P = .040), whereas it increased significantly with the advent of systemic metastasis (P = .027). In vitro, either serum from breast cancer patients or a medium originated from coculture experiments of MCF-7 cells and lymphocytes from healthy women stimulated heparanase expression in normal lymphocytes. The results suggest that there is a tumor-inducing effect on heparanase expression by lymphocytes present in the PBMCs of breast cancer patients, which depends, in turn, on the interaction between a tumor and normal lymphocytes

Profile of MicroRNAs Associated with Death Due to Disease Progression in Metastatic Papillary Thyroid Carcinoma Patients

Papillary thyroid carcinoma (PTC) is the most common neoplasm of the endocrine system and has an excellent long-term prognosis, with low rates of distant metastatic disease. Although infrequent, there are cases of deaths directly related to PTC, especially in patients with metastatic disease, and the factors that could be associated with this unfavorable outcome remain a major challenge in clinical practice. Recently, research into genetic factors associated with PTC has gained ground, especially mutations in the TERT promoter and BRAF gene. However, the role of microRNAs remains poorly studied, especially in those patients who have an unfavorable outcome at follow-up. This paper aims to evaluate molecular markers related to the different pathological processes of PTC, as well as the histological characteristics of the neoplasm, and to compare this profile with prognosis and death from the disease using an analysis of patients treated for metastatic disease in a single tertiary cancer center. Evaluation of microRNA expression in paraffin-embedded tumor specimens was carried out by quantitative PCR using the TaqMan® Low Density Array (TLDA) system. Metastatic patients who died from progression of PTC had higher expressions of miR-101-3p, miR-17-5p, and miR-191-5p when compared to patients with stable metastatic disease. These findings are of great importance but should be considered as preliminary because of the small sample