GRIM-19 Expression and Function in Human Gliomas.

OBJECTIVE: We determined whether the expression of GRIM-19 is correlated with pathologic types and malignant grades in gliomas, and determined the function of GRIM-19 in human gliomas. METHODS: Tumor tissues were isolated and frozen at -80 just after surgery. The tissues consisted of normal brain tissue (4), astrocytomas (2), anaplastic astrocytomas (2), oligodendrogliomas (13), anaplastic oligodendrogliomas (11), and glioblastomas (16). To profile tumor-related genes, we applied RNA differential display using a Genefishing DEG kit, and validated the tumor-related genes by reverse transcription polymerase chain reaction (RT-PCR). A human glioblastoma cell line (U343MG-A) was used for the GRIM-19 functional studies. The morphologic and cytoskeletal changes were examined via light and confocal microscopy. The migratory and invasive abilities were investigated by the simple scratch technique and Matrigel assay. The antiproliferative activity was determined by thiazolyl blue Tetrazolium bromide (MTT) assay and FACS analysis. RESULTS: Based on RT-PCR analysis, the expression of GRIM-19 was higher in astrocytic tumors than oligodendroglial tumors. The expression of GRIM-19 was higher in high-grade tumors than low-grade tumors or normal brain tissue; glioblastomas showed the highest expression. After transfection of GRIM-19 into U343MG-A, the morphology of the sense-transfection cells became larger and more spindly. The antisense-transfection cells became smaller and rounder compared with wild type U343MG-A. The MTT assay showed that the sense-transfection cells were more sensitive to the combination of interferon-beta and retinoic acid than U343MG-A cells or antisense-transfection cells; the anti-proliferative activity was related to apoptosis. CONCLUSION: GRIM-19 may be one of the gene profiles which regulate cell death via apoptosis in human gliomas.

Anticancer activity of PEGylated matrix metalloproteinase cleavable peptide-conjugated adriamycin against malignant glioma cells.

Although matrix metalloproteinases (MMPs) play a crucial role in the invasion and growth of malignant gliomas, their increased activity in tumor environment can be used as a specific target for chemotherapy. We investigated whether polymer-drug conjugates formed via MMP-cleavable peptide linkages could provide MMP-responsive tumor targeting and cytotoxicity for malignant glioma cells. One end of an MMP-cleavable peptide was attached to the end of methoxy polyethylene glycol (MPEG) while the other end was attached to adriamycin (ADR). The release of drugs in the presence of conditioned media of U87MG cells was investigated. The cytotoxicities of the MMP-cleavable MPEG-peptide-ADR (PPA) conjugates and non-cleavable MPEG-ADR (PA) conjugates were investigated using U87MG cells. The (1)H nuclear magnetic resonance (NMR) spectra confirmed the conjugation of the two ends of the peptide to the ends of MPEG and ADR, respectively. Gelatin zymography showed that MMP-2 was strongly expressed in the media of U87MG cells. The PA conjugate did not release ADR either in the phosphate buffered saline (PBS) or conditioned media of U87MG cells. The PPA conjugate released ADR in the presence of the conditioned media of U87MG cells, but not in PBS only. In the cytotoxicity test using U87MG cells, ADR and PPA conjugate showed similar anti-proliferative activities, while the cytotoxicity of PA conjugate was lower than that of ADR. Considering that the cytotoxicity of the PPA conjugate was similar to that of ADR, MMP-cleavable polymer-drug conjugates can be used as targeting carriers for the purpose of inhibiting the proliferation of malignant glioma cells.

The effect of hyaluronic Acid on the invasiveness of malignant glioma cells : comparison of invasion potential at hyaluronic Acid hydrogel and matrigel.

OBJECTIVE: Hyaluronidase (HAse), a degrading enzyme of hyaluronic acid (HA), is highly expressed in patients with malignant glioma. The purpose of this study was to verify whether HAse is related to the invasion of glioma cells. We also investigated if glioma cells with higher mobility in 2-dimensioal (2-D) method have also higher mobility at 3-dimensional (3-D) environment. METHODS: Malignant glioma cell lines (U87MG, U251MG, U343MG-A, and U373MG) were used, and their HAse expressions were evaluated by HA zymography. The migration ability was evaluated by simple scratch technique. The invasiveness of each cell lines was evaluated by Matrigel invasion assay and HA hydrogel invasion assay. In HA hydrogel invasion assay, colonies larger than 150 microm were regarded as positive ones and counted. Statistical analysis of migration ability and invasion properties of each cell lines was performed using t-test. RESULTS: In scratch test to examine migration ability of each cell lines, U87MG cells were most motile than others, and U343MG-A least motile. The HAse was expressed in U251MG and U343MG-A cell lines. However, U87MG and U373MG cell lines did not express HAse activity. In Matrigel invasion assay, the cell lines expressing HAse (U251MG and U343MG-A) were more invasive in the presence of HA than HAse deficient cell lines (U87MG and U373MG). In HA hydrogel invasion assay, the HAse-expressing cell lines formed colonies more invasively than HAse-deficient ones. CONCLUSION: Malignant Glioma cells expressing HAse were more invasive than HAse-deficient ones in 3-dimensional environment. Therefore, it might be suggested that invasion of malignant gliomas is suppressed by inhibition of HAse expression or HA secretion. Additionally, the ability of 2-D migration and 3-D invasion might not be always coincident to each other in malignant glioma cells.

Role of galectin-1 in migration and invasion of human glioblastoma multiforme cell lines.

OBJECT: Galectin-1 is highly expressed in motile cell lines. The authors investigated whether galectin-1 actually modulates the migration and invasion of human glioblastoma multiforme (GBM) cell lines, and whether its expression with respect to invasion and prognosis is attributable to certain glioma subgroups. METHODS: In the human GBM cell lines U343MG-A, U87MG, and U87MG-10', the RNA differential display was evaluated using Genefishing technology. The results were validated by reverse transcription polymerase chain reaction and Northern blot analysis to detect possible genetic changes as the determining factors for the motility of the malignant glioma. The migration and invasion abilities were investigated in human GBM cell lines and galectin-1 transfectant using an in vitro brain slice invasion model and a simple scratch technique. The morphological and cytoskeletal (such as the development of actin and vimentin) changes were examined under light and confocal microscopy. Galectin-1 expression was assessed on immunohistochemical tests and Western blot analysis. RESULTS: Endogenous galectin-1 expression in the human GBM cell lines was statistically correlated with migratory abilities and invasiveness. The U87-G-AS cells became more round than the U87MG cells and lacked lamellipodia. On immunohistochemical staining, galectin-1 expression was increased in higher-grade glioma subgroups (p = 0.027). CONCLUSIONS: Diffuse gliomas demonstrated higher expression levels than pilocytic astrocytoma in the Western blot. Galectin-1 appears to modulate migration and invasion in human glioma cell lines and may play a role in tumor progression and invasiveness in human gliomas.

Cisplatin-incorporated hyaluronic acid nanoparticles based on ion-complex formation.

The aim of this study is to prepare cisplatin-incorporated nanoparticles based on ion complex formation between hyaluronic acid (HA) and cisplatin for antitumor drug delivery. To prepare nanoparticles using HA, bulk HA was degraded by hyaluronidases (HAses). Cisplatin-incorporated HA nanoparticles were prepared by mixing cisplatin with an aqueous solution of HA and then the nanoparticle solution was dialyzed to remove trace elements. Since glioma tumor cell lines are able to secrete HAse, extracts from U343MG and U87MG cell lines were used to test the release of cisplatin from the nanoparticles. The morphological observation of the cisplatin-incorporated nanoparticles showed that they had spherical shapes with a particle size around 100-200 nm. The loading efficiency of cisplatin in the nanoparticles was about 67-81% (w/w) and cisplatin was continuously released from the nanoparticles for 4 days. Especially, the release rate of cisplatin from the nanoparticles increased when HAse was added to the release medium. In the results of the HA zymography, the U343MG cell line secreted HAse, while the U87MG cell line did not. When the extracts from U343MG were added to the release medium, the release rate of cisplatin was slightly increased, while the extracts from U87MG did not significantly affect the release rate of cisplatin. In conclusion, cisplatin-incorporated nanoparticles have sufficiently small particle sizes to use as a drug targeting system. The release of cisplatin from the nanoparticles was responsive to the secretion of HAse. These nanoparticles are suitable vehicles for an antitumor drug targeting system.

Prevention of postoperative subdural fluid collections following transcortical transventricular approach.

BACKGROUND: Subdural fluid collections appear in about 39% of patients after the removal of intra- and paraventricular tumors. This extracerebral fluid collection requires surgical intervention when progressive fluid accumulation takes place. The authors retrospectively and prospectively studied the efficacy of gelfoam and fibrin adhesive in closing cortical and ependymal defects after intraventricular and/or paraventricular lesion resection to prevent the development of SFCs. METHODS: From 1999 to 2004, we used gelfoam and fibrin adhesive on the cortical and ependymal defects of 28 patients who underwent the resection of intraventricular and/or paraventricular lesions via the transcortical approach associated with the communicated ventricle. We investigated the percentage of symptomatic and asymptomatic SFC. RESULTS: The patients median age was 59.5 years (range, 30-76 years), and the male/female ratio was 16:12. A frontal approach was performed in 18 patients, an occipital approach in 2, a parietal approach in 4, and a temporal approach in 4. The incidence of SFCs was 7% (2 patients). Of the 2 patients with SFCs, 1 required temporary drainage. The other patient was asymptomatic, and the SFCs were spontaneously absorbed 2 months later. CONCLUSIONS: The use of gelfoam and fibrin adhesive to seal cortical and ependymal defects after a transcortical procedure might be a viable method of preventing the development of SFC.

Cystic vestibular schwannomas: a possible role of matrix metalloproteinase-2 in cyst development and unfavorable surgical outcome.

OBJECT: The authors evaluated the clinical manifestations and surgical results in patients with cystic vestibular schwannoma (VS), and investigated the matrix metalloproteinase (MMP) expression of the cyst fluid and wall in an attempt to elucidate the pathogenesis and characteristics of this disease. METHODS: The clinical and neuroimaging features, perioperative findings, and surgical outcomes in 24 cases of cystic VS and 82 cases of solid VS, all of which were treated using the suboccipital approach, were retrospectively compared. To evaluate the role of MMP in cystic VS, gelatin zymography and immunohistochemical studies of the cyst fluid, wall, and solid portion were performed in nine cases of this disease. The mean duration of symptoms was shorter (14.0 months compared with 26.1 months; p = 0.04) and the mean size of the tumor was larger (43.8 mm compared with 34.2 mm; p = 0.048) in the cystic than the solid VS group. Although gross-total resection was easier to accomplish in this group (100% compared with 84.1%), adhesion to the facial nerve was more frequent (62.5% compared with 48.8%; p = 0.042). On gelatin zymography studies, MMP-2 expression was ubiquitously observed in all cyst fluids. Immunohistochemical analysis of the cyst wall showed that MMP-2 was apparently localized to the tumor cells on the luminal inner surface, adjacent to the cyst cavity. CONCLUSIONS: Resection of cystic VS is complicated by severe adhesion of the tumor capsule to the facial nerve and the large size of the lesion. The authors believe that MMP-2 may be involved in the pathogenesis of cyst formation or in its enlargement and may aggravate adhesion to the facial nerve, either by promoting the enlargement of the tumor or engendering the degradation of the tumor-nerve barrier proteolytically.

Solitary intracranial subdural osteoma: intraoperative findings and primary anastomosis of an involved cortical vein.

We report a patient with a intracranial subdural osteoma with a large cortical vein passing through the subdural calcified mass. A 60-year-old man presented with an approximately 3-year history of persistent headache. Computerized tomography (CT) scanning showed a homogeneous high-density nodule attached to the inner surface of the right frontal skull. Intraoperatively, the hard mass was found to be located in the intradural subarachnoid space. A large cortical vein passed through the subdural mass and was anastomosed in an end-to-end fashion after the excision of the segment involved by the tumor. The histopathologic examination showed lamellated bony trabeculae lined by osteoblasts and the underlying dura was uninvolved by the tumor cells.

Screening for motility-associated genes in malignant astrocytoma cell lines.

The most characteristic feature of a malignant astrocytoma is its early and extensive infiltration into adjacent parenchymal structures. We focused on detecting the possible expression changes as the determining factors for malignant astrocytoma's motile ability. We confirmed that four of 39 genes showed different expression on DD-PCR by RT-PCR and Northern blot analysis. These findings suggest that the genes identified may be important for determining high motility in astrocytoma cell lines. These findings may help us understand the molecular invasion mechanism in astrocytomas.

Polyion complex micelles composed of all-trans retinoic acid and poly (ethylene glycol)-grafted-chitosan.

The goal of this study is to develop novel types of polyion complex micelles for the drug delivery to brain tumor. Methoxy poly(ethylene glycol) (mPEG)-grafted chitosan (CP) was synthesized in order to make polymeric micelles encapsulating all-trans retinoic acid (ATRA) based on polyion complex formation. Polyion complex micelles were found to have spherical shapes with sizes of about 50 approximately 200 nm. The loading efficiency of micelle was higher than 80% (w/w) for all formulations. 1H nuclear magnetic resonance (NMR) spectra confirmed the formation of polymeric micelles. The CP graft copolymer and ATRA have distinguishing peaks in their 1H NMR spectra. The specific peaks of ATRA disappeared in D2O or DMSO while it appeared at mixtures of D2O/DMSO, indicating that ATRA and chitosan formed ion complex inner-core. In the cell cytotoxicity study using U87MG cells in vitro, polyion complex micelles showed similar cytotoxicity to that of free ATRA. A migration test was performed to investigate the inhibition of tumor cell invasion in vitro. The results suggested that the polyion complex micelles was more effective at inhibiting tumor cell migration than free ATRA.

Dumbbell-shaped middle cranial fossa meningioma with interdural cavernous sinus extension: report of two cases with complete removal.

BACKGROUND: Surgery for meningiomas involving the cavernous sinus remains controversial. Interdural cavernous sinus is called the lateral dural wall in the cavernous sinus, which is composed of two layers, the outer dural layer and the inner membranous layer. We encountered two cases of dumbbell-shaped middle cranial fossa meningioma with interdural cavernous sinus extension, which were successfully removed by surgical means. CASE DESCRIPTION: A 57-year-old woman presented with headache and decreased visual acuity. Neurological assessment was normal. Computed tomography and magnetic resonance imaging showed the presence of a dumbbell-shaped, smooth-contoured, well-enhanced mass in the right mesial temporal area. The lateral wall of the cavernous sinus was exposed via frontotemporal craniotomy and the tumor originating in the lateral wall was totally removed. A 41-year-old man presented with seizure attacks and drowsy mental status. Magnetic resonance imaging showed the presence of a multilobulated, well-enhanced mass in the left parasellar area. The tumor was totally resected via a transsylvian temporopolar approach. The mass originated from tentorial edge and extended into the cavernous sinus by dural penetration. CONCLUSION: Middle cranial fossa meningioma with interdural cavernous sinus extension can be removed more easily than other tumors with intracavernous sinus extension and, consequently, can be safely resected without any resulting cranial nerve deficit.

Expression of Nm23 in gliomas and its effect on migration and invasion in vitro.

BACKGROUND: Despite the well-documented importance of Nm23 in the control of metastasis, there is currently a paucity of data regarding the role of this gene family in the control of glioma invasion. MATERIALS AND METHODS: Nm23-H1 expression in gliomas was assessed via immunohistochemistry, Western blot, RT-PCR and Northern blot analyses. The migration and invasion ability were also investigated in primary glioma culture cells, human glioma cell lines and nm23-H1 transfectant, using an in vitro brain slice invasion model and a simple scratch technique. RESULTS: Although no significant correlations were detected between nm23-H1 expression and pathological grade, the endogenous nm23-H1 expression in gliomas was found to be inversely correlated with their migratory abilities. Additionally, the nm23-H1 transfectant resulted in a reduction of approximately 45% of the migratory ability and suppressed the invasiveness of the parental cell line. CONCLUSION: Our overall findings suggest that nm23-H1 may play an important role in the suppression of glioma invasion and migration.