ABSTRACT
The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth factor ß1 (TGF-ß1) and p53 in C6 cells of rat glioma in vitro. MATERIALS AND METHODS: FRBCS was obtained from suspensions of fetal rat brain cells on the 14th (E14) day of gestation. C6 glioma cells were cultured for 48 h in the presence of FRBCS or FRBCS + anti-TGF-ß1 monoclonal antibody. Immunocytochemical staining for TGF-ß1 and p53 was performed. RESULTS: The proportion of TGF-ß1-immunopositive tumor cells in C6 glioma cultures was statistically significantly higher than in the control cell cultures of normal fetal rat brain. FRBCS reduced the proportion of TGF-ß1-immunopositive tumor cells and increased the proportion of p53-immunopositive cells in C6 glioma cultures. In cells cultured with FRBCS + anti-TGF-ß1 monoclonal antibody, the above effects of FRBCS were abrogated. CONCLUSION: The obtained results suggest that TGF-ß1 seems to be responsible for decrease in TGF-ß1 expression and increase in p53 expression in C6 glioma cells treated with FRBCS.
Subject(s)
Brain Neoplasms/metabolism , Glioma/metabolism , Transforming Growth Factor beta1/biosynthesis , Animals , Cell Line, Tumor , RatsABSTRACT
The impact of rat neurogenic progenitor cells supernatant (RPNS) on the cytotoxic function of lymphocytes in rats under conditions of physiological norm and experimentally modeled tumor (brain glioma strain 101.8) was studied. The research was carried out in animals with inoculated tumor without RPNS injection and with different regimes of RPNS injection (thrice repeated from 5th to 10th day after glioma inoculation as well as 1 week and 1 month before tumor inoculation). Comparison groups included rats without glioma who triple injected with RPNS; and intact animals (control). RPNS was received from suspension of neurogenic progenitor cells (NPC) of rat brain on 14th day of gestation and injected intraperitoneally (0,12 mg per animal). Cytotoxic function of lymphocytes of experimental rats was evaluated in MTT-colorimetric test with allogeneic glioma cells. RPNS administration increased the cytotoxic activity of lymphocytes in vitro tests with allogeneic tumor cells in intact animals (to 37-38%) as well as in rats with glioma (to 11-22%). Under the RPNS influence the life expectancy and median survival of tumor-bearing animals increased (an average of 3-4 days). RPNS input modes such as triple injection from 5th to 10th day after glioma inoculation and 1 week before inoculation were the most effective. Thus, indirect tumor-inhibiting effect under intraperitoneal. RPNS administration in rats with glioma is demonstrated, which is obviously due to increased efficiency of cytotoxic function of immune cells of animals with inoculated tumor under the influence of the factors produced by NPC.
Subject(s)
Antineoplastic Agents/pharmacology , Brain Neoplasms/drug therapy , Culture Media, Conditioned/pharmacology , Glioma/drug therapy , Immunologic Factors/pharmacology , Neoplasms, Experimental/drug therapy , T-Lymphocytes, Cytotoxic/drug effects , Animals , Brain/drug effects , Brain/immunology , Brain/pathology , Brain Neoplasms/immunology , Brain Neoplasms/mortality , Brain Neoplasms/pathology , Cytotoxicity, Immunologic/drug effects , Drug Administration Schedule , Glioma/immunology , Glioma/mortality , Glioma/pathology , Injections, Intraperitoneal , Male , Neoplasms, Experimental/immunology , Neoplasms, Experimental/mortality , Neoplasms, Experimental/pathology , Neural Stem Cells/cytology , Neural Stem Cells/immunology , Neural Stem Cells/metabolism , Primary Cell Culture , Rats , Survival Analysis , T-Lymphocytes, Cytotoxic/immunology , T-Lymphocytes, Cytotoxic/pathologyABSTRACT
UNLABELLED: The aim of the work was to investigate the antitumor efficacy of allogeneic tumor vaccine (ATV) modified with rat progenitor neural cell supernatant (RPNS) in rats with glioma 101.8. MATERIALS AND METHODS: The study was performed on 74 white random-bred rats. ATV was developed on the basis of glioma 101.8 cell suspension modified with RPNS (0.02 or 0.10 mg/ml). RPNS was prepared from suspension made from whole rat brain tissue on 14(th) (E14) day of gestation. Model of brain glioma 101.8 was reproduced by intracerebral injection of glioma 101.8 cell suspension. ATV was injected intraperitoneally in a volume of 0.2 ml per animal once on the 10(th) day after tumor transplantation. For survival analysis Kaplan - Mayer multiple assessments method was used. Cytotoxic activity of rat lymphocytes (effector cells) was evaluated in MTT-colorimetric test by determining the state of mitochondrial dehydrogenase enzymes in target cells (allogeneic glioma 101.8 cells). RESULTS: Intraperitoneal administration of ATV modified with 0.10 mg/ml RPNS significantly increased mean survival time and median survival of glioma-bearing rats compared with unvaccinated group (MST (19.9 ± 2.4), 21.4 days; versus MST (14.6 ± 2.8); 14 days; p = 0.0002, Gehan's - Wilcoxon test). Intraperitoneal administration of ATV modified with 0.10 mg/ml RPNS resulted in increased cytotoxic activity of immune cells of rats with glioma in vitro compared with this index in unvaccinated group (p = 0.026, U-Mann - Whitney test). CONCLUSION: Antitumor effect of vaccination with RPNS-modified ATV is realized via increased cytotoxic activity of immune cells, what could be used for further optimization of whole tumor cell vaccine.
