氯法齐明增强免疫治疗的抗胶质瘤效果

Method of obtaining TMZ-resistant GB cells included treatment of T98G glioblastoma cells with 150 μmol/l TMZ. To confirm resistance to TMZ, MTT assay was performed according to the manufacturer's protocol. Untreated cells were used as a control group. C6 glioma cells were stereotactically implanted into the brain of Wistar rats and irradiated (24 Gy) in combination with oral administration of TMZ (20 mg/ kg) and clofazimine (CFZ) (30 mg/kg). This was followed by subsequent immunotherapy including tumor cell and dendritic cell vaccines. Neurovisualisation, immunocytochemical and immunohistochemical assays were used and animals' survival was analyzed with Kaplan-Meier estimator.

T98G resistant glioblastoma cell line is characterized by immunoreactive β-catenin, CD133, CD44, and N-cadherin as compared to the control cell line. The IC 50 of clofazimine for T98G glioblastoma cell line is 38.3 ± 4,1 μmol/l, for C6 rat glioma cell line is 37,6 ± 3,2 μmol/l. Clofazimine enhanced the cytotoxic activity of temozolomide, paclitaxel, and carboplatin in cancer cells of T98G line as compared to the control group. The cytotoxic effect of lomustine and carboplatin against T98G resistant glioblastoma cells was also enhanced by Clofazimine. Tumor cell vaccine (TCV) and dendritic cell vaccine (DCV) in combination with clofazimine produces a stronger anti-tumor immune response in C6 glioma. This is evident with development of local inflammatory reaction with higher content of interleukin 1β and 18 in serum, as well as greater level of IBA1+, CD68 + in pro-inflammatory microglia of neoplastic tissues. Combined use of DCV and clofazimine results in higher survival rates in experimental animals (- 90 ± 7 days against 45 ± 5 days) in the treated group with chemoradiation therapy (CRT).

Combination of clofazimine and immunotherapy enhances anti-glioma effect of TMZ in an in vivo model experiment.