Experimental validation of deuterium oxide—mediated antitumoral activity as it relates to apoptosis in murine malignant astrocytoma cells

2002 ◽  
Vol 96 (5) ◽  
pp. 900-908 ◽  
Author(s):  
Takeshi Uemura ◽  
Kouzo Moritake ◽  
Yasuhiko Akiyama ◽  
Yoriyoshi Kimura ◽  
Takashi Shingu ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Deuterium Oxide ◽  
Caspase Activation ◽  
Malignant Astrocytoma ◽  
Content Type ◽  
Activation Pathway ◽  
Astrocytoma Cells ◽  
M Phase ◽  
Induced Apoptosis ◽  
Fine Print

Object. Deuterium oxide (D2O), or heavy water, affects a variety of biological activities different from those of water. The authors examined the antitumoral effect of D2O on brain neoplasms and demonstrated D2O-mediated cytotoxicity by using a Rous sarcoma virus—induced murine malignant astrocytoma cell line, RSVM. The mechanism of the observed cytotoxicity may involve D2O-induced apoptosis and cell-cycle modulation. Methods. The authors performed an assay with methylthiazol tetrazolium bromide and a trypan blue dye exclusion test to confirm in vitro D2O-mediated cytotoxicity for RSVM cells. At D2O concentrations of 10 to 50%, the cytotoxic effect was dose and time dependent. Flow cytometry analysis revealed programmed cell death (apoptosis) and the accumulation of RSVM cells during the G2/M phase. By applying the terminal deoxynucleotidyl transferase—mediated deoxyuridine triphosphate nick-end labeling method, fluorescein isothiocyanate—annexin V and propidium iodide double staining, and caspase-family protease activity analysis, the authors demonstrated both DNA fragmentation and enhancement of caspase activity after a 48-hour treatment with D2O, thus indicating that D2O induces apoptosis in RSVM cells. Apoptotic DNA fragmentation was completely abolished by the caspase inhibitor Z-VAD-FMK (benzyloxycarbonil-Val-Ala-Aps-fluoromethylketone). The findings indicate that the caspase activation pathway may be involved in D2O-induced apoptosis. Conclusions. The authors found that D2O is cytotoxic to malignant astrocytoma cells. The mechanism of D2O-mediated cytotoxicity involved the induction of apoptosis and cell accumulation during the G2/M phase. This D2O-induced apoptosis is modulated through the caspase activation pathway.

Malignant astrocytoma following radiotherapy of a craniopharyngioma

1978 ◽  
Vol 48 (4) ◽  
pp. 622-627 ◽  
Author(s):  
Richard L. Sogg ◽  
Sarah S. Donaldson ◽  
Craig H. Yorke
Keyword(s):  
Experimental Evidence ◽  
Temporal Lobe ◽  
Malignant Astrocytoma ◽  
Content Type ◽  
Suprasellar Region ◽  
The Right ◽  
Fine Print

✓ A 9-year-old schoolgirl received 6007 rads to the suprasellar region for craniopharyngioma. Five years later, a malignant astrocytoma developed in the right temporal lobe. We cite clinical and experimental evidence to support our suspicion that the glioma may have been induced by radiation.

Serum proteolytic activity during the growth of C6 astrocytoma

1992 ◽  
Vol 77 (4) ◽  
pp. 595-600 ◽  
Author(s):  
Indrasen S. Vaithilingam ◽  
Warren McDonald ◽  
Noel K. Brown ◽  
Eric Stroude ◽  
Robert A. Cook ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Type Iv Collagen ◽  
Collagenase Activity ◽  
Type Iv Collagenase ◽  
Content Type ◽  
Type Iv ◽  
Astrocytoma Cells ◽  
Vessel Remodeling ◽  
C6 Astrocytoma ◽  
Fine Print

✓ Tumor growth is dependent on the ability of neoplastic cells to induce angiogenesis. Blood-vessel remodeling requires the reconstruction of the nonfibrous proteins and type IV collagen components of the basement membrane. This study has assessed the influence of the growth of C6 astrocytoma cells in the rat spheroid implantation model on serum general protease and type IV collagenase activity. The results demonstrate that general protease activity increased in serum, reaching maximum values on Day 6 and Day 13 following spheroid implantation, and that type IV collagenase activity increased in serum, obtaining maximum values on Day 8 and Day 15. The measurement of serum proteolytic activity may be of value in the detection of recurrent tumors.

Establishment and characterization of a novel malignant astrocytoma cell line derived from a tumor removed in a patient with neurofibromatosis Type 1

2001 ◽  
Vol 94 (2) ◽  
pp. 301-308 ◽  
Author(s):  
Masanori Kurimoto ◽  
Yutaka Hirashima ◽  
Tsuneaki Ogiichi ◽  
Hideo Hamada ◽  
Hironaga Kamiyama ◽  
...  
Keyword(s):  
Cell Line ◽  
Neurofibromatosis Type 1 ◽  
Proliferative Activity ◽  
Neurofibromatosis Type ◽  
Malignant Astrocytoma ◽  
Content Type ◽  
Astrocytoma Cell Line ◽  
Astrocytoma Cell ◽  
Fine Print

Object. Patients with neurofibromatosis Type 1 (NF1) have a predisposition to development of a variety of benign and malignant tumors including neurofibromas, astrocytomas, pheochromocytomas, and malignant peripheral nerve sheath tumors. The availability of an astrocytoma cell line derived from NF1 would be useful in studies in which sporadic astrocytomas could be compared with NF1-derived astrocytomas. In this article the authors describe a novel astrocytoma cell line, TM-31, that they established from a tumor removed in a 42-year-old woman with NF1. Methods. The TM-31 cell line was prepared from a surgical specimen of malignant astrocytoma and was serially subcultured over 250 times throughout a 6-year period without showing any sign of cell senescence. Immunocytochemical analyses demonstrated that TM-31 cells are negative for glial fibrillary acidic protein but positive for vimentin and S-100 protein. The TM-31 cells display little neurofibromin expression when subjected to immunoblotting, indicating that there is an NF1 gene mutation. Polymerase chain reaction—single-strand conformational polymorphism analysis revealed that TM-31 cells harbor a p53 point mutation in exon 7, codon 238. Chemosensitivity testing of TM-31 cells revealed a resistance to 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea, although they are sensitive to cisplatin and etoposide. In addition, TM-31 cells displayed no morphological differentiation after all-transretinoic acid and dibutyryl cyclic adenosine monophosphate treatments. Pharmacological inhibition of farnesyltransferase of the Ras oncoprotein led to decreased proliferative activity and inhibition of anchorage-independent growth of TM-31 cells in soft agar. Conclusions. The TM-31 cell line is an immortalized astrocytoma cell line derived from a tumor obtained in a patient with NF1. Ras activation may be the major event of proliferative activity and of the transformed phenotype of TM-31 cells, and the farnesyltransferase inhibitor may be potentially important as a novel antiproliferative therapy for NF1-derived astrocytomas.

Resistance of leukemic cells to 2-chlorodeoxyadenosine is due to a lack of calcium-dependent cytochrome c release

Blood ◽  
2002 ◽  
Vol 99 (2) ◽  
pp. 655-663 ◽  
Author(s):  
Joya Chandra ◽  
Emma Mansson ◽  
Vladimir Gogvadze ◽  
Scott H. Kaufmann ◽  
Freidoun Albertioni ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Dna Fragmentation ◽  
Cell Lines ◽  
Resistant Cell ◽  
Leukemic Cells ◽  
Caspase Activation ◽  
Cytochrome C Release ◽  
Deoxyguanosine Kinase ◽  
Induced Apoptosis ◽  
Resistant Cells

Abstract The purine nucleoside 2-chlorodeoxyadenosine (CdA) is often used in leukemia therapy. Its efficacy, however, is compromised by the emergence of resistant cells. In the present study, 3 CdA-resistant cell lines were generated and characterized. Their ability to accumulate 2-chloroadenosine triphosphate (CdATP) varied, reflecting differences in activities of deoxycytidine kinase (dCK) and deoxyguanosine kinase (dGK). Nonetheless, the selected lines were uniformly resistant to CdA-induced apoptosis, as assessed by caspase activation and DNA fragmentation. In contrast, cytosols from resistant cells were capable of robust caspase activation when incubated in the presence of cytochrome c and dATP. Moreover, replacement of dATP with CdATP also resulted in caspase activation in the parental and some of the resistant cell lines. Strikingly, CdA-induced decreases in mitochondrial transmembrane potential and release of cytochrome c from mitochondria were observed in the parental cells but not in any resistant lines. The lack of cytochrome c release correlated with an increased ability of mitochondria from resistant cells to sequester free Ca2+. Consistent with this enhanced Ca2+buffering capacity, an early increase in cytosolic Ca2+after CdA treatment of parental cells but not resistant cells was detected. Furthermore, CdA-resistant cells were selectively cross-resistant to thapsigargin but not to staurosporine- or Fas-induced apoptosis. In addition, CdA-induced caspase-3 activation and DNA fragmentation were inhibited by the Ca2+ chelator BAPTA-AM in sensitive cells. Taken together, the data indicate that the mechanism of resistance to CdA may be dictated by changes in Ca2+-sensitive mitochondrial events.

Enhancement of antitumor immune response in glioma models in mice by genetically modified dendritic cells pulsed with Semliki Forest virus—mediated complementary DNA

2001 ◽  
Vol 94 (3) ◽  
pp. 474-481 ◽  
Author(s):  
Ryuya Yamanaka ◽  
Susan A. Zullo ◽  
Ryuichi Tanaka ◽  
Michael Blaese ◽  
Kleanthis G. Xanthopoulos
Keyword(s):  
Dendritic Cells ◽  
Cancer Vaccines ◽  
Semliki Forest Virus ◽  
Glioma Cell Line ◽  
Tumor Lysate ◽  
Tumor Challenge ◽  
Content Type ◽  
Induced Apoptosis ◽  
Phosphate Buffered Saline ◽  
Fine Print

Object. The aim of this study was to further investigate dendritic cell (DC)—based immunotherapy for malignant glioma to improve its therapeutic efficacy. Methods. Dendritic cells were isolated from the bone marrow and pulsed with phosphate-buffered saline, tumor RNA, tumor lysate, Semliki Forest virus (SFV)-LacZ, SFV-mediated B16 complementary (c)DNA, or SFV-mediated 203 glioma cDNA, respectively, to treat mice bearing tumors of the 203 glioma cell line. The results indicated that preimmunization with DCs pulsed with the same type of cDNA as in the tumor by a self-replicating RNA vector (that is, SFV) protected mice from tumor challenge, and that therapeutic immunization prolonged the survival of mice with established tumors. The SFV induced apoptosis in DCs and their death facilitated the uptake of apoptotic cells by other DCs, thus providing a potential mechanism for enhanced immunogenicity. Conclusions. Therapy with DCs that have been pulsed with SFV-mediated tumor cDNA may be an excellent procedure for the development of new cancer vaccines.

Aberrant nuclear factor-κB activity and its participation in the growth of human malignant astrocytoma

2002 ◽  
Vol 96 (5) ◽  
pp. 909-917 ◽  
Author(s):  
Shoichi Nagai ◽  
Kazuo Washiyama ◽  
Masanori Kurimoto ◽  
Akira Takaku ◽  
Shunro Endo ◽  
...  
Keyword(s):  
Cell Growth ◽  
Cell Lines ◽  
Antisense Oligodeoxynucleotide ◽  
Nuclear Factor ◽  
High Grade ◽  
Malignant Astrocytoma ◽  
Content Type ◽  
High Grade Astrocytoma ◽  
Astrocytoma Cell ◽  
Fine Print

Object. It has been suggested that nuclear factor (NF)-κB, a pleiotropic transcription factor, controls cell proliferation. The authors examined NF-κB activity and its participation in the growth of human malignant astrocytoma. Methods. The authors examined NF-κB activity in human malignant astrocytoma cell lines and high-grade astrocytoma tissues by using electrophoretic mobility shift assays and immunohistochemical studies, respectively. In addition, messenger (m)RNA expression of p50 and RelA, which are representative subunits of NF-κB, and IκBα, which is a representative inhibitory protein of NF-κB, were analyzed using Northern blot hybridization in the astrocytoma cell lines. Furthermore, alterations in DNA synthesis and cell growth in the astrocytoma cell lines were examined after inhibition of NF-κB activity by RelA antisense oligodeoxynucleotide. The authors found NF-κB activity in all astrocytoma cell lines and high-grade astrocytoma tissues that were examined, but not in the fetal astrocyte strain or in normal cerebral tissue. Expression of p50, RelA, and IκBα mRNA was found in the fetal astrocyte strain and normal adult brain tissue, in addition to the astrocytoma cell lines. The relative levels of expression of these mRNAs were similar among these cell lines, the cell strain, and normal tissue. The RelA antisense oligodeoxynucleotide specifically reduced the levels of RelA mRNA expression and NF-κB activity in the astrocytoma cell lines, thus significantly inhibiting their DNA synthesis and cell growth. Conclusions. Human malignant astrocytoma cells have aberrant NF-κB activity, which promotes their growth. This activity is not associated with aberrant expression of p50 and RelA.

Marked enhancement of antitumor immune responses in mouse brain tumor models by genetically modified dendritic cells producing Semliki Forest virus—mediated interleukin-12

2002 ◽  
Vol 97 (3) ◽  
pp. 611-618 ◽  
Author(s):  
Ryuya Yamanaka ◽  
Susan A. Zullo ◽  
Jay Ramsey ◽  
Naoki Yajima ◽  
Naoto Tsuchiya ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Brain Tumor ◽  
Tumor Antigens ◽  
Semliki Forest Virus ◽  
Tumor Regression ◽  
Interleukin 12 ◽  
Content Type ◽  
Retrovirus Vector ◽  
Induced Apoptosis ◽  
Fine Print

Object. The authors evaluated dendritic cell (DC)—based immunotherapy for malignant brain tumor to improve its therapeutic efficacy. Methods. Dendritic cells were isolated from bone marrow and pulsed with phosphate-buffered saline, Semliki Forest virus (SFV)—LacZ, retrovirus vector GCsap—interleukin (IL)-12, and SFV—IL-12, respectively, to treat mice bearing brain tumors of the B16 cell line. The results indicated that therapeutic immunization with DCs pulsed with SFV—IL-12 prolonged the survival of mice with established tumors. Semliki Forest virus induced apoptosis in DCs, which in turn facilitated the uptake of apoptotic cells by other DCs, thus providing a potential mechanism for enhanced immunogenicity. Conclusions. Therapy with DCs that have been pulsed with SFV-mediated IL-12 may be an excellent step in the development of new cancer vaccines. Intratumorally injected DCs that have been transiently transduced with IL-12 do not require pulsing of a source of tumor antigens to induce tumor regression.

Inhibition of DNA repair for sensitizing resistant glioma cells to temozolomide

2003 ◽  
Vol 99 (6) ◽  
pp. 1047-1052 ◽  
Author(s):  
Takao Kanzawa ◽  
Joshua Bedwell ◽  
Yasuko Kondo ◽  
Seiji Kondo ◽  
Isabelle M. Germano
Keyword(s):  
Cell Cycle ◽  
Dna Repair ◽  
Cell Viability ◽  
Cell Cycle Arrest ◽  
Glioma Cells ◽  
Glioblastoma Cell ◽  
Content Type ◽  
Cycle Arrest ◽  
M Phase ◽  
Fine Print

Object. Temozolomide (TMZ) is a DNA alkylating agent currently used as adjuvant treatment for anaplastic astrocytomas. Its use in managing glioblastoma multiforme has been halted because of the lack of therapeutic effects due to cell resistance. Note that O6-alkylguanine—DNA alkyltranferase (AGT) is a DNA repair enzyme that limits the efficacy of TMZ. In this study the authors investigated the ability of O6-benzylguanine (BG), an AGT inhibitor, to sensitize a glioblastoma cell line resistant to TMZ. Methods. The effects of TMZ alone (100 µg) and after exposure to BG (50 µg) were assessed in two glioblastoma cell lines, U373-MG and T98G, respectively, sensitive and resistant to TMZ. Cell viability was assessed using trypan blue; cell cycle analysis by fluorescence-activated cell sorter; and apoptosis and autophagy by terminal deoxynucleotidyl transferase—mediated deoxyuridine triphosphate nick-end labeling (TUNEL) and acridine orange staining, respectively. Furthermore, the involvement of an autophagy marker, microtubule-associated light chain 3 (LC3), was assessed. Temozolomide suppressed the growth of and caused cell cycle arrest in the G2—M phase of U373-MG cells but not T98G cells. Exposure to BG prior to TMZ resulted in a significant decrease in cell viability as well as cell cycle arrest in the G2—M phase in T98G cells (p < 0.05). Although apoptosis was not detected on TUNEL staining, programmed cell death Type II (autophagy) was detected after exposure to BG and TMZ in T98G cells. Conclusions. These results indicate that inhibition of AGT by BG can render previously resistant glioma cells sensitive to TMZ treatment. The mechanism of cell demise following BG-TMZ treatment seems to be autophagy and not apoptosis. Combination therapy involving TMZ and an AGT inhibitor may be an effective strategy to treat resistant gliomas.

Prognostic value of magnetic resonance imaging—guided stereotactic biopsy in the evaluation of recurrent malignant astrocytoma compared with a lesion due to radiation effect

2003 ◽  
Vol 98 (1) ◽  
pp. 14-20 ◽  
Author(s):  
Matthew J. McGirt ◽  
Ketan R. Bulsara ◽  
Thomas J. Cummings ◽  
Kent C. New ◽  
Kenneth M. Little ◽  
...  
Keyword(s):  
Magnetic Resonance ◽  
Malignant Glioma ◽  
Prognostic Value ◽  
Stereotactic Biopsy ◽  
Radiation Effect ◽  
Recurrent Glioma ◽  
Recurrent Malignant Glioma ◽  
Malignant Astrocytoma ◽  
Content Type ◽  
Fine Print

Object. The prognostic value of differentiating between recurrent malignant glioma and a lesion due to radiation effect by performing stereotactic biopsy has not been assessed. Thus, this study was undertaken to determine such value. Methods. Between 1995 and 2001, 114 patients underwent magnetic resonance (MR) imaging—guided stereotactic biopsy to differentiate lesions caused by a recurrence of malignant astrocytoma and by radiation effect. All patients had previously undergone tumor resection (World Health Organization Grade III or IV) followed by radiotherapy. Disease diagnosis based on biopsy and patient characteristics were assessed as predictors of survival according to results of a multivariate Cox regression analysis. The diagnosis determined with the aid of biopsy was compared with that established during a subsequent resection in 26 patients. Survival following stereotactic biopsy was markedly increased in patients suffering from radiation effect compared with those harboring recurrent malignant glioma (p < 0.0001). In patients with radiation effect on biopsy, an increasing patient age (p < 0.05), having had two compared with one prior resection (p < 0.05), and a decreasing time from radiotherapy to biopsy (p < 0.001) were factors associated with decreased survival. Nevertheless, in patients with biopsy-defined radiation effect at second progression or with an age younger than 50 years the survival rate remained higher than that in patients with recurrent tumor on biopsy (p < 0.01). A biopsy-based diagnosis of radiation effect obtained less than 5 months after radiotherapy was not associated with an increased rate of patient survival compared with a diagnosis of recurrent malignant glioma on biopsy (p = 0.286). Eighty-six percent of lesions initially determined to be due to radiation effect on biopsy fewer than 5 months after radiotherapy were characterized as recurrent glioma by a mean of 11 months later. In contrast, only 25% of lesions initially diagnosed as attributable to radiation effect on biopsy more than 5 months after radiotherapy were classified as recurrent glioma a mean of 12 months later (p < 0.05). Conclusions. With the aid of stereotactic biopsy the authors demonstrated prognostic significance in differentiating recurrent malignant astrocytoma from a lesion due to radiation effect in patients presenting more than 5 months after having undergone radiotherapy. In patients who presented earlier than 5 months after radiotherapy, radiation effect on biopsy was not associated with an improved rate of survival compared with that in patients harboring recurrent malignant astrocytoma.

Development of cerebellar malignant astrocytoma at site of a medulloblastoma treated 11 years earlier

1978 ◽  
Vol 49 (3) ◽  
pp. 445-449 ◽  
Author(s):  
Enrique Klériga ◽  
Joanna Hollenberg Sher ◽  
Sanath-Kumar Nallainathan ◽  
Sherman C. Stein ◽  
Michael Sacher
Keyword(s):  
Cerebellar Hemisphere ◽  
Malignant Astrocytoma ◽  
Content Type ◽  
Left Cerebellar Hemisphere ◽  
Desmoplastic Medulloblastoma ◽  
Second Tumor ◽  
Fine Print

✓ A child treated for a desmoplastic medulloblastoma of the left cerebellar hemisphere at the age of 10 months developed a malignant astrocytoma in the same site 11 years later. Theories of origin of the second tumor, particularly in relation to concepts of the genesis of medulloblastoma in general, are discussed.

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