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Objective To study changes in the expression levels of OX-62, OX-6 and CD86 of mononuclear cells and the related chemotatic factor macrophage inflammatory protein-1α (MIP-1α) in the livers of rats with Walker-256 tumors treated with radiofrequency ablation (RFA) and to elucidate the influence of RFA on differentiation and migration of liver dendritic cells(DCs).Methods Walker-256 liver tumors were induced in 60 SpragueDawley rats by implanting tumor particles. These rats were randomly divided equally into three groups from which liver tissue around the local area of the tumor was sampled at 7 d and 14 d after RFA. The mononuclear liver cells were separated with Ficoll density gradient centrifugation. The expression levels of OX-62, OX-6 and CD86 in the mononuclear cells were analyzed with flow cytometry. The expression level of MIP-1α in the liver tissue was detected by enzyme-linked immunosorbent assay (ELISA). Results The average expression of OX-6 in the control rats was 15.29 ±4.59% and those 7 d and 14 d after RFA were 34.20±11.62% and 39.18±9.14% respectively. The difference between the two RFA groups and the control group was statistically significant. The average expression rate of OX-62 in the control rats was 18.91±4.58% and those 7 d and 14 d after RFA were 24.49±4.45% and 22.77 ± 3.50% respectively. The difference between the 7 d group and the control group was significant. The average expression rate of CD86 in the control rats was 66.29±17.69% and those 7 d and 14 d after RFA were 55.29±10.69% and 55.93±12.64% respectively. These differences between both RFA groups and the controls group were not significant. The average expression level of MIP-1 α around the tumors was 232.92±54.58 pg/ml in the controls and 499.38±15.14 pg/ml and 495.90±9.94 pg/ml 7d and 14 d after RFA respectively. These differences from the controls were both statistically significant. Conclusion The expression of MIP-1α around the tumors was elevated after RFA, which could promote the migration of DCs. The changes in the expression of OX-62, OX-6 and CD86 also could reflect increased DC differentiation, which could improve local antigen-presenting capacity to a certain extent and recovery of host anti-tumor immune response.
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Objective To study the change of spleen Dendritic cells in normal rats treated by radio-frequency ablation(RFA). Methods Eighteen healthy SD rats were separated into group 1 week after RFA with 6 rats,group 2 week after RFA with 6 rats and control group with 6 rats. Spleen tissue were taken out respectively before RFA, 1 week after RFA and 2 weeks after RFA. The number and the phenotype of Dendritic cells in spleen were analyzed with flowcytometry. Results Pathologyical examination after RFA showed the characteristic that coagulation necrosis and cellular degeneration and granulation tissue forming appeared from target center to peripheral of the target. (10. 36±3. 21) % of normal rat mononuclear cells in spleen express OX-62, the ratio became (18. 03±5. 7) % 1 week after RFA and (12. 63±8. 0) % 2 weeks after RFA, the difference between group 1 week after RFA and control group was marked. (76. 33±7. 86) % of normal rat mononuclear cells in spleen express OX-6,the ratio became (78.33±7.25)% 1 week after RFA and (86. 04±7. 25) % 2 weeks after RFA, the difference between group 2 weeks after RFA and control group was marked. (63. 06±8. 77) % of normal rat mononuclear cells in spleen express CD86,the ratio was (55. 74±14. 49)% 1 week after RFA and (63.49±11.81)% 2 weeks after RFA,the difference between groups 1 week or 2 weeks after RFA and control group was not marked. Conclusions RFA can increasing the number of precursor Dendritic cells migrating from peripheral blood to spleen, and those cells may furtherly differentiate or maturate, which may be contributed to improve the ability delivery of body to antigen to a certain extent.
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BACKROUND:Previous studies have confirmed that radiofrequency ablation(RFA)has not only efficiently killed tumor cells,but also improved immune suppression of organism.Antigen presenting cells play an important role during anti-tumor immune reaction.Dendritic cells are most powerful antigen presenting cells.OBJECTIVE:To study the influence of RFA on OX-62,OX-6 and CD86 expression of rat peripheral blood mononuclear cells.DESIGN,TIME AND SETTING:The randomized controlled animal observation was performed at the School of Oncology of Peking University from June 2005 to March 2006.MATERIALS:A total of 12 Sprague Dawley rats were equally and randomly assigned into control and 1 -week radiofrequency groups.METHODS:2 mL peripheral blood was extracted from rats of the control group following rats were sacrificed.The left lobe of rat liver was exposed,inserted with acicular electrodes at tilted position.The acicular electrodes were spread out.The action time was 4 minutes.After RFA,the rats were given anti-infective therapy.2 mL peripheral blood was taken out after they were put to death 1 week after RFA.MAIN OUTCOME MEASURES:The following parameters were measured:pathomorphological observation on the liver of normal rats after RFA;OX-62,OX-6 and CD86 expression of peripheral blood mononuclear cells before and after RFA.RESULTS:Pathologyical examination after RFA showed the characteristic that coagulation necrosis and cellular degeneration and granulation tissue forming appeared from target center to peripheral of the target.Positive expression rate of OX-62 in rat peripheral blood in the 1-week radiofrequency group[(0.70±0.16)%]was significantly higher than in the control group [(0.34±0.08)%,P ＜0.05].No significant difference in positive expression rate of OX-6 and CD86 in the peripheral blood mononuclear cells between both groups(P＞0.05).CONCLUSION:RFA can promote the increased number of precursor Dendritic cells in rat peripheral blood,which may be contributed to improve the ability to angtigen presentation during immune response.Dai Wei-de,Doctor,Associate chief physician,Department of Ultrasound,Beijing Hospital of Ministry of Public Health,Beijing 100730,China dai.weide@126.com
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Objective To discuss the change of serum TGF-β1 and IL-10 in peripheral blood of rats with liver tumor treated by RFA.Methods Thirty experimental liver tumor model of SD rats were prepared by implantation of tumor particles.These rats were randomly divided equally into three groups including 1 week after RFA,2 week after RFA and control group.Peripheral blood of control group,group 1 week after RFA and group 2 week after RFA was taken out respectively without RFA,1 week and 2 week after RFA.The mononuclear cells of peripheral blood were separated by Ficoll density gradient centrifugation.The expression level of IL-10 in peripheral blood was analyzed with flowcytometry.Serum TGF-β1 were dectected by ELISA.Results The serum expression level of TGF-β1 of control group was(6.61±0.12)μg/L,and that of group 1 week and 2 week after RFA were respectively(5.63±0.46)μg/L and(5.53±0.56)μg/L.There was statistical significance for the difference between control group and group 1 week or group 2 week after RFA.The serum expression level of IL-10 of control gorup was 95.92±2.31,and that of group 1 week and 2 week after RFA were respectively 89.71±5.44 and 87.67±11.11.There was statistical significance for the difference between control group and group 1 week after RFA.Conclusions RFA can destroy the tumor tissues in situ and relief immune suppression by IL-10,TGF-β1 secreted by tumor tissue.RFA can improve differentiation and maturation of dendritic cell in local area of tumor and promate ability of antigen-presentation of body.
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Objective To study the mechanisms of inhibiting proliferous effect of human breast cancer cell line treated with genistein in vitro. Methods Two human breast cancer cell lines (MCF 7 and MDA MB 231) were cultured in vitro , proliferation of them was measured with MTT method, cell morphology was observed with Giemsa stain, cell cycle and apoptosis rate were measured with flow cytometry and apoptosis was detected in situ. Results Genistein markedly inhibited proliferation of MCF 7 and MDA MB 231 cell line. Typical image of apoptosis was observed in MCF 7 cell line treated with genistein in Giemsa stain:cell condensation, blebs formation on cell surface and pieces of condensed chromatin were scattered along nuclear envelope. The results detected with flow cytometry showed G 1 subpeak before G 1 phase peak (apoptosis peak). Apoptosis rate increased with time of genistein treatment. However, MDA MB 231 cell line treated with genistein did not show apoptosis and apoptosis peak, and quantity of cells at G 2 M phase obviously increased.Conclusion Genistein inhibites proliferation of human breast cancer cell line in vitro through inducing apoptosis or arresting cell at G 2 M phase.\;[