Generation and measurement of low-temperature plasma for cancer therapy: a historical review.

This review provides a description of the historical background of the development of biological applications of low-temperature plasmas. The generation of plasma, methods and devices, plasma sources, and measurements of plasma properties, such as electron dynamics and chemical species generation in both gaseous and aqueous phases, were assessed. Currently, direct irradiation methods for plasma discharges contacting biological surfaces, such as the skin and teeth, are related to plasma biological interactions. Indirect methods using plasma-treated liquids are based on plasma-liquid interactions. The use of these two methods is rapidly increasing in preclinical studies and cancer therapy. The authors address the prospects for further developments in cancer therapeutic applications by understanding the interactions between the plasma and living organisms.

Mechanism of introduction of exogenous genes into cultured cells using DEAE-dextran-MMA graft copolymer as non-viral gene carrier.

Comparative investigations were carried out regarding the efficiency of introduction of exogenous genes into cultured cells using a cationic polysaccharide DEAE-dextran-MMA (methyl methacrylate ester) graft copolymer (2-diethylaminoethyl-dextran-methyl methacrylate graft copolymer; DDMC) as a nonviral carrier for gene introduction. The results confirmed that the gene introduction efficiency was improved with DDMC relative to DEAE-dextran. Comparative investigations were carried out using various concentrations of DDMC and DNA in the introduction of DNA encoding luciferase (pGL3 control vector; Promega) into COS-7 cells derived from African green monkey kidney cells. The complex formation reaction is thought to be directly proportional to the transformation rate, but the complex formation reaction between DDMC and DNA is significantly influenced by hydrophobic bonding strength along with hydrogen bonding strength and Coulomb forces due to the hydrophobicity of the grafted MMA sections. It is thought that the reaction is a Michaelis-Menten type complex formation reaction described by the following equation: Complex amount = K1 (DNA concentration)(DDMC concentration). In support of this equation, it was confirmed that the amount of formed complex was proportional to the RLU value.

A phase I clinical trial of interferon-beta gene therapy for high-grade glioma: novel findings from gene expression profiling and autopsy.

BACKGROUND: High-grade gliomas are highly lethal neoplasms representing approximately 20% of all intracranial tumors. Cationic liposome-mediated interferon-beta (IFN-beta) gene transfer has been found to induce regression of experimental glioma. We have previously performed a pilot clinical trial to evaluate the safety and effectiveness of this IFN-beta gene therapy in five patients with high-grade glioma. Two patients showed more than 50% reduction while others had stable disease 10 weeks after treatment initiation. METHODS: To identify alterations in gene expression in brain tumors 2 weeks after the gene therapy trial, we used a microarray technology and Gene Ontology analysis. The results were validated by patients' clinical course and findings of histology and autopsy. RESULTS AND CONCLUSIONS: Using hierarchical clustering and principal component analysis, five series of gene therapy trials were classified according to the response to IFN-beta gene therapy. Significant changes in gene expression related to immunoresponse and apoptosis were observed. Moreover, novel patterns of altered gene expression, such as inhibition of neovascularization, were identified, suggesting the involvement of pathways reported previously as not involved. Autopsy and histological examinations revealed dramatic changes in the tumor tissues after therapy in all patients. Many tumor cells showed necrotic changes, and immunohistochemistry identified numerous CD8-positive lymphocytes and macrophages infiltrating the tumor and surrounding tissues; these were probably the effects of therapy. Simultaneously, CD34-immunoreactive vessels were notably decreased in the vector-injected brain. This study facilitates the understanding of the antitumor mechanism and helps identify candidate target molecules for new approaches. However, additional clinical trials are warranted.

Significant antitumor activity of cationic multilamellar liposomes containing human interferon-beta gene in combination with 5-fluorouracil against human renal cell carcinoma.

Immunotherapy is one of the most effective treatments against metastatic renal cell carcinoma (RCC). However, the response rate is not high. Therefore, more effective therapies are necessary for patients with metastatic RCC. We previously reported on the significant antitumor activity of cationic multilamellar liposome containing human interferon-beta (huIFN-beta) gene (IAB-1) against RCC. We then examined the antitumor effect of IAB-1 in combination with anticancer drugs against RCC. The cytotoxicity of IAB-1 alone, and in combination with anticancer drugs, cisplatin, adriamycin, 5-fluorouracil, gemcitabine, paclitaxel and irinotecan hydrochloride against the human RCC cell line NC65 was examined by the colorimetric method using tetrazolium salt. For the in vivo study, we used NC65 cells inoculated into the severe combined immunodeficiency mouse. The results showed that the in vitro combination therapy with IAB-1 and 5-FU was more cytotoxic than IAB-1 alone. However, synergistic cytotoxicity was not observed when combined with IAB-1 and other anticancer drugs. NC65 tumors transfected with IAB-1 in mice were smaller than those receiving an injection of empty liposome or the recombinant huIFN-beta protein. Treatment with IAB-1 in combination with 5-FU resulted in significant anticancer activity. IAB-1 enhanced the activity of thymidine phosphorylase (TP), which converts 5-FU to the active metabolite, FdUMP. In contrast, IAB-1 decreased the activity of thymidylate synthase (TS), which is a target enzyme of 5-FU. In conclusion, these findings indicate that a combination of IAB-1 and 5-FU may have enhanced antitumor activity against human RCC, suggesting its potential clinical application. The mechanism of enhanced cytotoxicity by combination therapy with IAB-1 and 5-FU may up-regulate TP activity and down-regulate TS activity.

A pilot study of human interferon beta gene therapy for patients with advanced melanoma by in vivo transduction using cationic liposomes.

BACKGROUND: Cationic liposomes containing the human interferon beta (HuIFNbeta) gene (IAB-1) was used for the clinical trial for glioma patients. HuIFNbeta gene therapy showed much higher anti-tumor activity compared with the administration of HuIFNbeta protein for melanoma. These results suggest that HuIFNbeta gene therapy is an attractive strategy for the treatment of melanoma. METHODS: Stage IV or III melanoma patients with cutaneous or subcutaneous metastatic lesions were enrolled in this pilot study. IAB-1 was dissolved by sterile PBS at a concentration of 30 microg DNA/ml and was injected into cutaneous or subcutaneous metastatic nodules three times a week for 2 weeks and the effect on the injected and non-injected metastatic lesions was evaluated. RESULTS: Clinical responses were as follows (five patients): mixed response (MR) and no change in each one patient, and progressive disease in three patients. In the MR patient, the IAB-1 injected lesion disappeared clinically and histopathologically and one-half of IAB-1 non-injected skin metastases were transiently inflamed and mostly regressed. In the responded non-injected lesions of this patient, histopathologically, infiltration of CD4 positive T cells was observed around the melanoma cells in the dermis, which expressed the HLA-Class II antigen. Adverse events due to this gene therapy were not recognized in any of the patients. CONCLUSIONS: The efficacy of this gene therapy was generally insufficient; however, some immunological responses were recognized in one patient. No adverse events were observed. HuIFNbeta gene therapy could be an attractive strategy for treatment of a variety of malignancies, including melanoma, though some modifications should be required.

[Molecular targeted therapy for renal cell carcinoma].

Renal cell carcinoma (RCC) accounts for approximately 2% of all cancer cases worldwide. Metastatic disease is often present at the time of diagnosis of RCC and its poor response to chemotherapy and radiotherapy causes poor prognosis. Immunotherapy is relatively effective for RCC, but the response rate is approximately 15-20%. Therefore, new therapeutic approaches are necessary for these patients with metastatic RCC. Recently, the mechanisms responsible for the growth of RCC have been clarified, and molecular targeted therapy has been developed. In this paper, we review the new molecular targeted therapeutic agents effective for RCC.

Supermolecular drug challenge to overcome drug resistance in cancer cells.

Overcoming multidrug resistance (MDR) of cancer cells can be accomplished using drug delivery systems in large-molecular-weight ATP-binding cassette transporters before entry into phagolysosomes and by particle-cell-surface interactions. However, these hypotheses do not address the intratumoral heterogeneity in cancer. Anti-MDR must be related to alterations of drug targets, expression of detoxification, as well as altered proliferation. In this study, it is shown that the excellent efficacy and sustainability of anti-MDR is due to a stable ES complex because of the allosteric facilities of artificial enzymes when they are used as supermolecular complexes. The allosteric effect of supermolecular drugs can be explained by the induced-fit model and can provide stable feedback control systems through the loop transfer function of the Hill equation.

Characteristics of DEAE-dextran-MMA graft copolymer as a nonviral gene carrier.

A stable and soapless latex of diethylaminoethyl-dextran-methyl methacrylate (DEAE-dextran-MMA) graft copolymer (DDMC) has been developed for nonviral gene delivery vectors that are possible to autoclave. DDMC relatively easily formed a polyion complex between DNA and DDMC by the hydrophobic force of graft poly(MMA) depending on its large positive entropy change (DeltaS). DDMC has been confirmed as having a high protection facility for DNase by DNase degradation test.Transfection activity was determined using the beta-galactosidase assay, and a higher value of 16 times or more was confirmed for the DDMC samples in comparison with one of the starting DEAE-dextran hydrochloride samples. The resulting DDMC, having an amphiphilic domain so as to form a polymer micelle, should become a stable latex with a hydrophilic-hydrophobic microseparated domain. The complex of DDMC and plasmid DNA may be formed on the spherical structure of the amphiphilic microseparated domain of DDMC and have a good affinity to the cell membrane. The infrared absorption spectrum shift to a high-energy direction at around 3450 cm(-1), because of the complexes between DNA and DDMC, may cause the formation of more compact structures, not only by a coulomb force between the phosphoric acid of DNA and the DEAE group of DEAE-dextran copolymer but also by a force from the multi-intermolecule hydrogen bond in the backbone polymer DEAE-dextran and a hydrophobic force from the graft poly(MMA) in DDMC. It is thus concluded that DNA condensation may possibly have a high transfection efficiency via DDMC. The high efficiency of this graft copolymer, which is sterilized by an autoclave, may thus make it a valuable tool for safe gene delivery.

Growth inhibition of human pancreatic cancer cells by human interferon-beta gene combined with gemcitabine.

We examined the anti-tumor effect of cationic multilamellar liposome containing human IFN-beta (huIFN-beta) gene against cultured human pancreatic cancer cells. We also evaluated the combined effect of huIFN-beta gene entrapped in liposomes and gemcitabine. Furthermore, we examined the anti-tumor mechanisms of the therapy, with emphasis on the Ras-related signal pathway. Three human pancreatic cancer cell lines (AsPc-1, MIAPaCa-2, and PANC-1) were used in this study. The growth inhibition together with the therapy were evaluated by WST-1 assay; the production of huIFN-beta protein was measured by ELISA; the cell cycle and apoptosis were analyzed using a FACScan flow cytometer; the protein levels of Son of sevenless (SOS-1) and Ras-GAP were measured by Western blotting; and the activation of Ras-GTP was evaluated by the immunoprecipitation method. As a result, we found that huIFN-beta gene entrapped in liposomes demonstrated a strong anti-tumor effect against human pancreatic cancer cells. The treatment that combined huIFN-beta gene entrapped in liposomes and gemcitabine was more effective than each treatment alone. Although gemcitabine remarkably reduced the level of SOS-1, the above combined therapy reduced the level of SOS-1 even more significantly. Both huIFN-beta gene entrapped in liposomes and the com-bination of huIFN-beta gene entrapped in liposomes and gemcitabine increased the level of Ras-GAP, and decreased the activity of Ras-GTP. These results suggest that this combination therapy can induce strong anti-tumor activity against human pancreatic cancer cells through the regulation of the Ras-related signal pathway.

Significant antitumoral activity of cationic multilamellar liposomes containing human IFN-beta gene against human renal cell carcinoma.

PURPOSE: Immunotherapy is the most effective treatment against metastatic renal cell carcinoma (RCC). However, the response rate is approximately 15%. More effective therapy is, therefore, needed for patients with metastatic RCC. We then examined the antitumor effect of cationic multilamellar liposome containing human IFN-beta (huIFN-beta) gene (IAB-1) against RCC. EXPERIMENTAL DESIGN: Concentrations of huIFN-beta protein were measured by ELISA. The cytotoxicity of IAB-1 against human RCC (NC65, ACHN, and freshly isolated RCC cells), prostate and bladder cancer cell lines, and renal proximal tubule endothelial cells (RPTEC5899) was examined by the colorimetric method using tetrazolium salt. Apoptosis was assessed by the acridine-orange staining. For in vivo study, we used NC65 cells inoculated into severe combined immunodeficiency mouse. RESULTS: The RCC cells treated with IAB-1 secreted significant amounts of huIFN-beta protein continuously. Drastic in vitro cytotoxic effect of IAB-1 against RCC was observed. In contrast, treatment with 1000 IU/ml recombinant huIFN-beta protein resulted in weak cytotoxicity. The cytotoxic effect against prostate and bladder cancer cell lines was less than that against RCC. Furthermore, no significant cytotoxicity was observed in RPTEC5899 cells. Apoptosis was observed in the cells treated with IAB-1, but recombinant huIFN-beta failed to induce apoptosis. The size of NC65 tumors transfected with IAB-1 in mice was significantly smaller than that receiving injection of empty liposome or recombinant huIFN-beta protein. CONCLUSION: These findings indicate that IAB-1 may have an antitumor activity against human RCC by inducing apoptosis, suggesting its potential clinical application for gene therapy against RCC.

[Gene therapy using cationic multilamellar liposomes containing human interferon-beta gene against renal cell carcinoma].

The anticancer activity of cationic multilamellar liposomes containing human IFN-beta gene (IAB-1) against renal cell carcinoma (RCC) was examined. Concentrations of IFN-beta protein were measured by an enzyme-linked immunosorbent assay. The cytotoxic activity of IAB-1 against RCC cells and normal renal proximal tubule endothelial cells (RPTEC5899) was examined by the microculture tetrazolium dye assay. For the in vivo study, the NC65 RCC cell line was inoculated into severe combined immunodeficiency mouse. The RCC cells treated with IAB-1 secreted significant amounts of IFN-beta protein. Significant in vitro cytotoxic activity of IAB-1 against RCC cells was observed. In contrast, treatment of RCC cells with recombinant IFN-beta protein resulted in less cytotoxicity. No significant cytotoxicity was seen in RPTEC5899 cells. Apoptosis was observed in RCC cells treated with IAB-1. The size of NC65 RCC cancers transfected with IAB-1 in mice was significantly smaller than that receiving injection of empty liposomes or recombinant IFN-beta protein. These findings show that IAB-1 may have significant antitumor activity against RCC, and suggest its potential clinical application for gene therapy against RCC.

Medicinal facilities to B16F10 melanoma cells for distant metastasis control with a supramolecular complex by DEAE-dextran-MMA copolymer/paclitaxel.

The resistance of cancer cells to chemotherapeutic drugs (MDR) is a major problem to be solved. A supramolecular DEAE-dextran-MMA copolymer (DDMC)/paclitaxel (PTX) complex was obtained by using PTX as the guest and DDMC as the host having 50-300 nm in diameter. The drug resistance of B16F10 melanoma cells to paclitaxel was observed, but there is no drug resistance of melanoma cells to the DDMC/PTX complex in vitro. The cell death rate was determined using Michaelis-Menten kinetics, as the DDMC/PTX complex promoted allosteric supramolecular reaction to tubulin. The DDMC/PTX complex showed a very superior anti-cancer activity to paclitaxel alone in vivo. The median survival time (MST) of the saline, PTX, DDMC/PTX4 (particle size, 50 nm), and DDMC/PTX5 (particle size, 290 nm) groups were 120 h (T/C, 1.0), 176 h (T/C, 1.46), 328 h (T/C, 2.73), and 280 h (T/C, 2.33), respectively. The supramolecular DDMC/PTX complex showed the twofold effectiveness of PTX alone (p < 0.036). Histochemical analysis indicated that the administration of DDMC/PTX complex decreased distant metastasis and increased the survival of mice. A mouse of DDMC/PTX4 group in vivo was almost curing after small dermatorrhagia owing to its anti-angiogenesis, and it will be the hemorrhagic necrotic symptom of tumor by the release of "tumor necrosis factor alpha (TNF-α)" cytokine. As the result, the medicinal action of the DDMC/PTX complex will suppress the tumor-associated action of M2 macrophages and will control the metastasis of cancer cells.

Human gene therapy for malignant gliomas (glioblastoma multiforme and anaplastic astrocytoma) by in vivo transduction with human interferon beta gene using cationic liposomes.

Transfer of interferon beta gene via cationic liposomes has been found to induce regression of experimental glioma. We performed a pilot clinical trial of safety and effectiveness of this interferon beta gene therapy in five patients with malignant glioma (glioblastoma multiforme or anaplastic astrocytoma). Transgene expression and antitumor activity were detected in four patients. Two patients showed a partial response (>50% tumor reduction) and two others had stable disease 10 weeks after beginning therapy. One patient could not be evaluated because of previous treatment with gamma-knife therapy. This study suggests the feasibility and safety of interferon beta gene therapy, which may become an important treatment option for patients with malignant glioma.

Cationic liposomes conjugation to recombinant adenoviral vectors containing herpes simplex virus thymidine kinase gene followed by ganciclovir treatment reduces viral antigenicity and maintains antitumor activity in mouse experimental glioma models.

Gene therapy using adenoviral (Ad) vector containing herpes simplex virus thymidine kinase (AxCAHSV-tk) followed by the administration of ganciclovir (GCV) has been a promising therapy for cancer including malignant gliomas. However, there remain numerous problems to overcome, such as the high immunogenicity and toxicity of Ad vector. To optimize the therapy, we investigated whether a conjugation of our original cationic liposomes and Ad vectors reduces viral antigenicity and maintains the antitumor activity in mouse experimental (subcutaneous and intracranial) glioma models. Our original liposomes consist of N-(alpha-trimethylammonioacetyl)-didodecyl-D-glutamate chloride, dilauroyl phosphatidylcholine, and dioleoyl phosphatidyl-ethanolamine in a molar ratio of 1:2:2. AxCAHSV-tk and GCV showed a remarkable inhibition of experimental glioma growth. The growth-inhibitory effect decreased in mice previously immunized with another Ad vector (AxCALacZ). In contrast, the conjugation of AxCAHSV-tk and liposomes did not diminish the growth-inhibitory effect. Furthermore, the conjugation reduced antigenicity for Ad vector in vivo. These findings suggest that suicide gene therapy, using a conjugation of AxCAHSV-tk and our liposomes, is a feasible approach for human cancer gene therapy, especially malignant gliomas.

Repeated cationic multilamellar liposome-mediated gene transfer enhanced transduction efficiency against murine melanoma cell lines.

We investigated whether repeated cationic multilamellar liposome-mediated gene transfers enhanced the transduction efficiency against murine melanoma cell lines and experimental subcutaneous melanoma. In the former, the murine melanoma cell line, B16F10, was transfected by our original cationic multilamellar liposomes containing pVLacZ, which express beta-galactosidase in eukaryotic cells. Cells were exposed to the liposomes in a single, double, or triple procedure during the cell logarithmic proliferative period. We then evaluated the transduction efficiency by X-gal staining and beta-galactosidase assay. The number of positive cells and level of beta-galactosidase activity were significantly increased by repeated exposures compared with a single one. Cells transfected by the fluorescently labeled cationic liposome containing pEGFP-C1 showed both an increased uptake of liposomes and an increased number of EGFP expression cells following repeated exposures. In the latter, murine subcutaneous melanomas, which were made by transplantation of B16F10 in C57BL6 mice, were transfected by same liposomes. Subcutaneous melanomas were exposed to the liposomes in a single, double, or triple procedure. We then evaluated the transduction efficiency by the beta-galactosidase assay. The level of beta-galactosidase activity was significantly increased by repeated exposures compared with a single one. The results indicate that repeated exposures to the liposomes enhanced the transduction efficiency toward murine melanoma cells and experimental subcutaneous melanoma, and may provide a basis for the repeated-exposure protocol for human trials.

Growth inhibition of subcutaneous mouse melanoma and induction of natural killer cells by liposome-mediated interferon-beta gene therapy.

In this study we investigated the antitumour effect and mechanism of action of cationic liposome-mediated murine interferon-beta (IFNbeta) gene therapy in mouse B16F1 melanoma cells in vitro and in vivo. Murine IFNbeta gene transfer by cationic liposome resulted in substantial growth inhibition of B16F1 melanoma cells in culture when compared with phosphate buffered saline or recombinant murine IFNbeta treatment, or lacZ control gene transfer. Use of video-enhanced contrast-differential interference contrast (VEC-DIC) microscopy revealed that liposomes containing the murine IFNbeta gene [lip(pSV2muIFNbeta)], but not recombinant murine IFNbeta, induced dramatic morphological changes that characterize apoptosis, including bleb formation, shrinkage of cells, nuclear condensation and 'ballooning', in approximately 30% of the cells treated. Intratumoral administration of lip(pSV2muIFNbeta) resulted in a 5.5-fold reduction in the mean volume of subcutaneous melanoma lesions in syngeneic mice 15 days after treatment and eradicated the tumour in 18% of the mice treated. Immunocytochemical analysis demonstrated that a larger number of natural killer (NK) cells infiltrated the tumour following lip(pSV2muIFNbeta) treatment than in controls. In vivo depletion of NK cells using the anti-asialoGM1 antibody reduced the efficacy of lip(pSV2muIFNbeta) treatment. Taken together, our data suggest that cationic liposome-mediated IFNbeta gene therapy could be effective against melanoma by directly inducing cell death and stimulating NK cells.

Improvement of transduction efficiency of recombinant adenovirus vector conjugated with cationic liposome for human oral squamous cell carcinoma cell lines.

Adenovirus (Ad) vectors are commonly used in gene therapy trials because of their efficiency in gene transfer. However, their use is limited by immune responses that reduce transgene expression and decrease the efficiency of repeated vector administration. In this study, the efficacy of gene transduction and the tumor-cell killing effect on four human oral (SAS, HSC-2, HSC-3, HSC-4) and one murine squamous cell carcinoma cell (SCC-7, a kind gift of Dr. M. Hiraoka, Kyoto University) lines in vitro with Ad vector conjugated with catioic liposome (Ad/SUV) was evaluated. Ad/SUV resulted in two to five-fold over higher transduction efficiency in four human and one murine cell lines in vitro than Ad vector alone. The optimal Ad-SUV ratio was determined as 10(6) pfu of Ad vector with 1 micromol SUV. Ad/SUV showed more tumor-cell killing effect than Ad vector alone. Furthermore, the shielding effects of Ad vector with Ad/SUV from neutralizing antibody were evaluated. We also found that Ad/SUV is less susceptible to inactivation by neutralizing antibodies in vitro. The efficacy of gene transduction with Ad vector was blocked more than 70% with neutralizing serum, while Ad/SUV retained approximately 50% of the control activity in vitro. On the basis of these results, the anti-tumor effect with suicide gene therapy using Ad/SUV in vivo was evaluated. Three injections of Ad/SUV showed the inhibition of tumor growth compared with control in vivo. Our results suggested that an enhanced anti-tumor effect on human oral squamous cell carcinoma would be obtained with repeated administrations of Ad/SUV.

Significant induction of apoptosis in renal cell carcinoma cells transfected with cationic multilamellar liposomes containing the human interferon-ß gene through activation of the intracellular type 1 interferon signal pathway.

We previously reported that cationic multilamellar liposome containing the human interferon-ß (huIFN-ß) gene (IAB-1) demonstrated significant cytotoxic effect in the NC65 human renal cell carcinoma (RCC) cell line. In this study, we investigated the molecular mechanisms of IAB-1-induced apoptosis and cytotoxicity in RCC cells. Remarkable in vitro cytotoxic and apoptosis-inducing effects of IAB-1 against NC65 cells were observed by a colorimetric method and TUNEL staining, respectively. In contrast, treatment of NC65 cells with exogenously added huIFN-ß protein induced low-level cytotoxicity without apoptosis. Neutralizing antibodies against huIFN-ß significantly suppressed the cytotoxic effect of huIFN-ß protein, but they were unable to block the effect of IAB-1. Cytotoxicity assays using transwell plates revealed that NC65 cells treated with IAB-1 did not secrete cytotoxic soluble factors other than IFN-ß. Substantial enhancement of interferon-stimulated response element (ISRE) activity of NC65 cells by IAB-1 was demonstrated by promoter reporter assays. In addition, immunofluorescence using confocal microscopy revealed the intracellular expression of IFN-ß and its receptor induced by IAB-1. The induction of c-Myc by IAB-1 was suggested by a cDNA macroarray and was confirmed by western blot analysis. These findings indicate that IAB-1 induces significant cytotoxicity and apoptosis in NC65 cells, possibly through enhanced ISRE activity, that is associated with increased intracellular localization of huIFN-ß and IFN-receptor. Our data support the potential clinical application of IAB-1 gene therapy for RCC resistant to IFN.

Clinical gene therapy for brain tumors. Liposomal delivery of anticancer molecule to glioma.

Liposomes are one of the most promising delivery systems for genes, proteins, and other biological molecules and they are expected to become a new therapeutic tool for the treatment of brain tumors, especially malignant gliomas. Until now, transfer of anticancer molecules using liposomes has been studied by a lot of investigators and it has been found to induce regression of experimental gliomas, resulting in establishing some original and effective therapies. Gene therapy using cationic liposomes is also one of them. Here we introduce the advanced medicine for brain tumors using liposomes containing some anticancer molecules (for example, gene, antibody, antisense, or magnetite), based on our basic and clinical research.