1-2-dielaidoylphosphatidylethanolamine and Glioma

To study the effects of alpha1,4Gal T antisense oligonucleotide mediated by lipofectin on human glioma cell line SWO-38.. SWO38 glioma cells were exposed to 10 micromol/L alpha1,4Gal T antisense oligonucleotide for 72 h by means of lipofectin transfection, and the growth inhibition of the cells was detected by colony-forming unit assay. Analysis of DNA fragmentation was performed with flow cytometry (FCM) and agarose gel eletrophoresis with Fas protein expression determined by FCM.. alpha1,4Gal T antisense oligonucleotide significantly inhibited the growth of glioma cells (P<0.01) and induced apoptosis in SWO-38 cells (P<0.05). Marked up-regulation of Fas protein expression was observed in response to the treatment (P<0.01).. alpha1,4Gal T antisense oligonucleotide can significantly inhibit SWO-38 cell proliferation and induce cellular apoptosis, the mechanism of which may involve up-regulated Fas expression.

Topics: Antineoplastic Agents; Cell Division; fas Receptor; Galactosyltransferases; Glioma; Humans; Oligonucleotides, Antisense; Phosphatidylethanolamines; Transfection; Tumor Cells, Cultured; Tumor Stem Cell Assay; Up-Regulation

To aim at demonstrating whether cationic liposome-mediated antisense c-myb oligonucleotide(LipoAON) can inhibit the growth of C6 glioma by intravenous injection.. Intracerebral C6 glioma cells were implanted into the left caudal nucleus of forty-eight male Wistar rats. There were four groups: LipoAON(n = 12), antisense c-myb oligonucleotide (AON; n = 12), cationic liposome (Lipo; n = 12), and normal saline (NS; n = 12). Six days after tumor implantation, the above-mentioned drugs were injected into the right femoral veins of the rats respectively. Two days later, the same drugs were injected into the left femoral veins. The appetite, motor and weight of every animal were closely observed during the whole experiment. Six rats of each group were respectively killed 4 days and 10 days after the end of administration. The weight change, pathologic examination and immunohistochemical analysis of c-myb expression of the tumor were completed.. In LipoAON group, the growth of the tumors was significantly inhibited in a short time after treatment and c-myb expression was down-regulated. But in the AON group and Lipo group, the growth of the tumors was not inhibited and c-myb expression was not down-regulated, compared with that in NS group. The inhibitory effect of LipoAON on the tumors rapidly declined with time and c-myb expression was again up-regulated.. 1. Cationic liposome (LipofectAMINE) as transfection vehicle makes c-myb easily penetrate BBTB and enter the tumor. The technique is simple, safe, highly effective for the transfection of c-mybAON; 2. LipoAON has marked inhibitory effect on the growth of C6 glioma. The AON technical method for inhibiting the expression of c-myb oncogene has a research perspective in the treatment of glioma; 3. The inhibitory effect of LipoAON on the growth of glioma declines with time. The question about how to make c-myb AON have highly effective, sustained and stable expression in the tumor still requires further research.

Topics: Animals; Brain Neoplasms; Cations; Cell Division; Genes, myb; Glioma; Liposomes; Male; Oligonucleotides, Antisense; Phosphatidylethanolamines; Proto-Oncogene Proteins c-myb; Rats; Rats, Wistar; Tumor Cells, Cultured

Topics: Animals; Base Sequence; Brain Neoplasms; Cell Division; Cell Nucleus; Drug Carriers; Glioma; Indicators and Reagents; Liposomes; Microtubule-Associated Proteins; Oligodeoxyribonucleotides, Antisense; Phosphatidylethanolamines; Rats; RNA-Binding Proteins; RNA, Messenger; Thionucleotides; Tumor Cells, Cultured

Epidermal growth factor receptor (EGFR) plays an important role in the progression of malignancy in gliomas. We studied the growth inhibition of the malignant glioma cell lines using an antisense EGFR oligodeoxynucleotide enveloped with Lipofectin. At a concentration of 5 microM of the antisense EGFR oligodeoxynucleotide enveloped with Lipofectin, the proliferation of three malignant glioma cell lines was significantly inhibited (p < 0.05) compared with that of the cells exposed to 5 microM sense EGFR oligodeoxynucleotide. The activity of the tyrosine kinase and the DNA synthesis was also significantly suppressed (p < 0.05). These findings show that the antisense EGFR oligodeoxynucleotide enveloped with Lipofectin has a possibility to become a useful gene therapy against malignant gliomas.

Topics: Brain Neoplasms; Cell Division; DNA, Neoplasm; Drug Carriers; ErbB Receptors; Genetic Therapy; Glioma; Humans; Liposomes; Oligodeoxyribonucleotides, Antisense; Phosphatidylethanolamines; Transfection; Tumor Cells, Cultured

Using a minipump combined with stereotaxic techniques allows continuous delivery of therapeutic genetic materials into the brain. We investigated the therapeutic efficacy of liposome-mediated HSVtk gene transfer of experimental brain F98 glioma followed by treatment with ganciclovir. A single injection of DNA-liposome complexes showed a therapeutically significant decrease in the tumor volume. Continuous intracerebral delivery of DNA-liposome complexes using an osmotic minipump led to complete tumor regression in 36.4% of the treated animals. The safety and toxicity of this gene delivery system were also assessed. No organ pathology was observed in the experimental animals. The continuous gene delivery system could be a useful means of achieving higher doses with less toxicity and without the need for frequent injections.

Topics: Animals; Antiviral Agents; Brain Neoplasms; Cell Survival; DNA, Viral; Ganciclovir; Gene Transfer Techniques; Genes, Reporter; Genetic Therapy; Glioma; Humans; Lac Operon; Lipids; Liposomes; Phosphatidylethanolamines; Quaternary Ammonium Compounds; Rats; Rats, Inbred F344; Simplexvirus; Stereotaxic Techniques; Thymidine Kinase

We recently have developed a unique cytoplasmic transient gene expression system based on cotransfection of target cells with bacteriophage T7 RNA polymerase (RNAP) and plasmid DNA vectors containing a T7 autogene. Because this T7 system is self-initiating, self-maintaining, and requires no cellular factors for transcription, it is therefore likely to function in any mammalian cell with any gene both in vitro and, more importantly, in vivo. In this study we demonstrate that the T7 DNA vector and T7 RNAP could be efficiently codelivered to cultured cells by lipofection. Different target genes were expressed by the T7 system in a wide variety of mammalian cells including several tumor cell lines. Gene expression could be detected in more than 30% of the cells of some tumor cell lines transiently transfected by the T7 vector. Average activity of the reporter enzyme (luciferase) expressed by a transfected cell was relatively constant regardless of the cell line used. When a T7-luciferase vector was directly injected into various tissues of mice without the use of liposomes, luciferase activity could be found in the injected liver, muscle, brain and tail connective tissues. The luciferase levels expressed by the T7 system were found to be up to 200-fold higher, depending upon the injected tissues, than levels achieved with a traditional nuclear gene expression vector. Direct tumor injection with a T7-beta-galactosidase (beta-gal) construct resulted in beta-gal gene expression in tumor cells near the injection sites. In addition, direct injection of the T7 system in mice did not generate detectable quantities of antibodies against the T7 RNAP. These results suggest that this gene expression system may be useful in many different medical applications such as cancer gene therapies and DNA vaccination, where transient but rapid and efficient gene expression is required.

Topics: Animals; Antibodies, Viral; Bacteriophage T7; beta-Galactosidase; Cation Exchange Resins; CHO Cells; Cricetinae; Cytoplasm; DNA-Directed RNA Polymerases; Fibrosarcoma; Gene Expression; Genes, Reporter; Genes, Viral; Genetic Therapy; Genetic Vectors; Glioma; Humans; L Cells; Lipids; Liver; Luciferases; Mice; Mice, Inbred BALB C; Mice, Nude; Molecular Sequence Data; Osteosarcoma; Phosphatidylethanolamines; Plasmids; Promoter Regions, Genetic; Rats; Transfection; Tumor Cells, Cultured; Viral Proteins

The Myc family proteins represented by c-Myc are thought to play a crucial role in cellular proliferation, differentiation, transformation, and apoptosis. In this study, we demonstrated the novel role for a Myc family protein in elicitation of immunogenic phenotypes in tumor cells. Injection of rat 9L or C6 glioma cells, together with the s-myc gene linked to the cytomegalovirus promoter, completely prevented formation of both brain tumors and s.c. tumors derived from the parental glioma cells. However, introduction of the s-myc gene had no inhibitory effect on development of B104-derived neuroblastoma. In addition, unlike the s-myc gene, injection of the c-myc or wild type p53 (wt-p53) gene together with glioma cells did not modulate the tumor immunogenicity and resulted in formation of gliomas in the animals. These findings suggest that s-Myc expression may stimulate the presentation of a tumor antigen common to 9L and C6 cells to T lymphocytes and augment the activity of the host immune system, resulting in prevention of glioma formation in vivo. This success in tumor eradication indicates the possibility of application of the s-myc gene for gene therapy of human brain tumors.

Topics: Animals; Brain Neoplasms; Genes, myc; Genes, p53; Genetic Therapy; Glioma; Hindlimb; Male; Neoplasm Transplantation; Neuroblastoma; Phosphatidylethanolamines; Proto-Oncogene Proteins c-myc; Rats; Rats, Inbred F344; Skin Neoplasms; Transfection; Tumor Cells, Cultured