| Objective:To investigate the effect of pseudomonas aeruginous MSHA (PA-MSHA) on cell invasive ability of C6 glioma cells in vitro; To study the growth regulation effect and potentiai mechanism of rat C6 glioma cell in body by PA-MSHA. Explore the mechanism of tumor cell growth inhibition action caused by the PA-MSHA in body. To explore new method of effectively postoperative adjuvant therary for glioma.Methods:Adopting Matrigel simulated reconstruction of the basement membrane, using Transwell chambers to establish the interaction model. The effect of various concentrations PA-MSHA monoclonal antibody on C6 glioma cells invasive ability in vitro was measured by Transwell chambe invasive models. To establish the SD rat C6 glioma allograft animal model, the weight of body between 180-200g. We conducted a non-blinded, randomized, controlled trial.40 rats with glioma were randomly assiganed to receive either 0.9% Nacl (control group, n=20) or the left by PA-MSHA vaccine (experimental group n=20). View the general state of the tumor-bearing mice after administration; Observe the value-added rate of tumor. After that we used HE and TEM to study the glioma and Flow Cytometry to investigate the Immune function before and after treatmeat.Results:1. Invasion assay the blank group, C6 glioma cells cell markedly enhanced invasive ability, a large number of cells through Matrigel, to reach the bottom of the cell membrane number is (13.65±2.18), C6 glioma cell under the action of PA-MSHA (1×10~9cfu/ml) invasive ability has been lowed, There are a number of cells through Matrigel, to reach the bottom of the cell membrane number is (30.10±1.45), C6 glioma cell under the action of 2×10~9cfu/ml invasive ability is not strong, a low number of cells through Matrigel, to reach the bottom of the cell membrane is (54.60±2.68). C6 glioma cells under the action of the PA-MSHA invasion ability as compared with compared groups, differences were significant (P＜0.001). 2. To establish the SD rat C6 glioma allograft animal modelC6 glioma cells used in this experiment,were adherent growth under conventional culture conditions and be determinated of dynamic before eath vaccination. The survival rate were always over 95%。The SD rat were be touched like the size of grain of rice after 8 days, and all the rats can be touched tumor after 10 days.3. View the general state of the tumor-bearing mice and observe the value-added rate of tumor after administration.The control group rats have less food and water than the experimental group after vaccination. Contract the experimental group, the control group showed decreased activity and body weakness, increase the incidence of symptoms such as limb paralysis by the time.The value-added rate of tumor was 26.4% in the experimental group and 100% in the control group (P＜0.05)4. We used HE and TEM to study the glioma.All the rats were resected the tumor under anesthesia in the 22 days after inoculation, The growth C6 glioma cells of the experimental group in lower density than the control group and some regions appear in liquefaction necrosis. by HE, and can see the smaller size of cell contraction, nuclear pyknosis deep-dyed blue morphological changes of apoptosis cells by TEM, showing early apoptotic changes.5. Flow cytometry to investigate the Immune function.Collected the serum at different time by the flow cytometry, The level of the IL-2,r-IFN and the TNF-a in the experimental group are higher than control group (P＜0.05),but the level of the IL-5 and IL-4 are lower than control group (P＜0.05) Conclusion:Pseudomonas aeruginosa (PA-MSHA) can positively regulate the cell proliferation and invasive ablity of C6 glioma, and provide more experimental bases for gliom treatment by PA-MSHA.In body experiments, PA-MSHA preparations can suppress the proliferation of C6 glioma cells. The mechanism may be related enhance Thl-cell function and reduce Th2-cell function. The immune functions, especially the cellular immunity are incressed significantly. It can also activate cytokine network in the cell line.
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