Effects of inducible glial fibrillary acidic protein on glioma cell motility and proliferation

El‐Obeid, Adila; Bongcam‐Rudloff, Erik; Westermark, Bengt; Nistér, Monica

doi:10.1002/(sici)1097-4547(20000415)60:2<245::aid-jnr14>3.0.co;2-1

Cited by 40 publications

(24 citation statements)

References 39 publications

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“…The outer proliferative zone did not express GFAP. This is consistent with reports that GFAP is rarely expressed in cycling cells in human glioma in situ [15]. GFAP expression was lost from the spheroid centre as an area of cell death developed.…”

Section: Glioma Spheroid Characterizationsupporting

confidence: 93%

The development of necrosis and apoptosis in glioma: experimental findings using spheroid culture systems*

Bell

Whittle

Walker

et al. 2001

Neuropathology Appl Neurobio

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Cell death in gliomas may occur either by apoptosis, or, in the case of high grade tumours, by necrosis, but questions remain as to the pathogenesis and relationship between these processes. The development of cell death was investigated in multicellular glioma spheroid cultures. Spheroids model the development of cell death due to diffusion gradients in a three-dimensional system without confounding influences of immune response, pressure gradients, etc. Spheroid cultures were established from four malignant glioma cell lines: U87, U373, MOG-G-CCM and A172; harvested from culture at weekly intervals and stained with Haematoxylin and Eosin (H&E), TdT-mediated dUTP-X nick end labelling (TUNEL) and by immunohistochemistry for vimentin, Glial Fibrillary Acidic Protein (GFAP) and Ki67. Annexin V flow cytometry and counts of apoptotic cells on H & E stained sections were performed to assess levels of apoptosis. Modes of cell death were also characterized by electron microscopy. Spatially separate zones of proliferation, differentiation and central cell death developed with increasing spheroid diameter. Central cell death developed at a predictable radius (300-400 microm) for each cell line. Ultrastructural examination showed this to be necrotic in type. Apoptosis was most reliably assayed by morphological counts using H & E. Basal levels of apoptosis were low (< 0.5%), but increased with increasing spheroid diameter (> 2% in U87). In particular, levels of apoptosis rose following development of central necrosis and apoptoses were most abundant in the peri-necrotic zone. There were quantitative differences in the levels of apoptosis and necrosis between glioma cell lines. The predictable onset of necrosis in the spheroids will allow us to investigate the pathogenesis of necrosis and events in prenecrotic cells. There is a relationship between the development of necrosis and apoptosis in this model and these processes can be separately assayed. Further in vitro and genetic studies will enable us to study these events and interactions in greater detail than is possible using other cell culture and in vivo systems.

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Section: Glioma Spheroid Characterizationsupporting

confidence: 93%

The development of necrosis and apoptosis in glioma: experimental findings using spheroid culture systems*

Bell

Whittle

Walker

et al. 2001

Neuropathology Appl Neurobio

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“…GFAP is the most widely used marker of astroglial cells, and its protein level appears to correlate inversely with the malignancy of gliomas (Deck et al, 1978;Eng and Rubinstein, 1978;Jacque et al, 1979;de Armond et al, 1980;Duffy, 1982;Duffy et al, 1982;Rutka et al, 1997). Consistent with these observations, high-level expression of the exogenously added GFAP gene in glioma cells was reported to reduce cell motility, invasiveness and proliferation (Rutka and Smith, 1993;Rutka et al, 1994Rutka et al, , 1997Murphy et al, 1998;Toda et al, 1999;Elobeid et al, 2000). Indeed, the transition from a GFAP-positive to a GFAP-negative astrocyte was often interpreted to represent transformation to a blas-tic, less mature, more aggressive state.…”

Section: Discussionmentioning

confidence: 87%

Downregulation of GFAP, TSP‐1, and p53 in human glioblastoma cell line, U373MG, by IE1 protein from human cytomegalovirus

Lee

Jeon

Kim

et al. 2005

Glia

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Human cytomegalovirus (HCMV) is a member of the b-herpesvirus family, which has tropism for glial cells. It was recently reported that HCMV might play important roles in the pathogenesis of malignant glioma. In this study, we investigated the effects of the HCMV IE1 protein on the gene expression profile in the human glioblastoma cell line, U373MG by employing cDNA microarray technology. Using DNA chips containing approximately 1,000 human cDNAs, RNA samples from U373MG cells stably expressing IE1 were compared with those from the control cells lacking IE1 cDNA. Fluorescence intensities of 13 genes were significantly decreased in IE1-expressing cells, while one gene was found to be upregulated. Among these 14 genes, we chose to work further on glial fibrillary acidic protein (GFAP), thrombospondin-1 (TSP-1), and p53, because of their previously known involvement in tumorigenesis. The mRNA levels of all these genes were found to be decreased in IE1-expressing glioblastoma cells by real-time quantitative reverse transcriptionpolymerase chain reaction (RT-PCR) as well as Northern blot analysis. The decreased expression of these genes was also observed at protein levels as measured by immunocytochemistry or fluorescence-activated cell sorting (FACS) analysis. Our data strongly suggested that HCMV IE1 could modulate the expression of cellular genes that might play important roles in the pathogenesis of glial tumors. '

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“…Moreover, a relationship between increased GFAP protein expression and suppression of glial tumor growth [30], inhibition of cellcycle progression, and decreased proliferation was previously reported [31]. The increased expression of GFAP in human glioblastoma cells was also related to reduced cell motility [32], maybe as a consequence of the interactions of this intermediate filament protein with actin and other cytoskeletal proteins.…”

Section: Discussionmentioning

confidence: 88%

Ukrain modulates glial fibrillary acidic protein, but not connexin 43 expression, and induces apoptosis in human cultured glioblastoma cells

et al. 2007

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Glioblastoma is a highly malignant tumor, characterized by an unfavorable prognosis even in response to multidisciplinary treatment strategies, owing to its high-invasive phenotype. Ukrain, a semisynthetic thiophosphoric acid derivative of the purified alkaloid chelidonine, has been used in the therapy of several solid tumors, but little is known about its effect on glioblastoma and, in general, about the molecular mechanisms responsible for its effects. In particular, we previously demonstrated that Ukrain modulates the expression of genes and proteins involved in tumor invasion, and here we investigate some unreported effects of Ukrain on human cultured glioblastoma cells. We used morphological and molecular biology methods to analyze the expression and the intracellular distribution pattern of glial fibrillary acidic protein, the expression of the gap junction protein connexin 43 and the apoptotic effect in human glioblastoma cells treated with 0.1, 1 and 10 micromol/l Ukrain for 72 h. After treatment with 10 micromol/l Ukrain, glial fibrillary acidic protein fluorescence increased and a higher number of cells displayed glial fibrillary acidic protein organized into a filamentous state. Western blot analysis of glial fibrillary acidic protein confirmed that Ukrain tended to upregulate the protein. Connexin 43 was not modulated by Ukrain both at the mRNA and at the protein level. Ukrain-induced apoptotic rate was 4.63, 10.9 and 28.9% after 0.1, 1 and 10 micromol/l Ukrain, respectively, likely mediated by cytochrome c release in the cytoplasm. Considered as a whole, these findings provide new information to complete the understanding of the mechanisms of Ukrain antitumor and chemopreventive effect, and support the possible potential of Ukrain for the therapy of brain tumors.

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Effects of inducible glial fibrillary acidic protein on glioma cell motility and proliferation

Cited by 40 publications

References 39 publications

The development of necrosis and apoptosis in glioma: experimental findings using spheroid culture systems*

The development of necrosis and apoptosis in glioma: experimental findings using spheroid culture systems*

Downregulation of GFAP, TSP‐1, and p53 in human glioblastoma cell line, U373MG, by IE1 protein from human cytomegalovirus

Ukrain modulates glial fibrillary acidic protein, but not connexin 43 expression, and induces apoptosis in human cultured glioblastoma cells

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