树突状细胞治疗大鼠肝癌的实验研究

摘要

目的通过负载癌抗原的树突状细胞（DC）治疗大鼠原发性肝癌(HCC)模型的实验，探讨其临床生物治疗的可行性。方法用大鼠HCC细胞株CBRH-7919细胞匀浆粗提物，刺激Wistar大鼠骨髓衍化的DC，按不同途径（静脉、腹腔、癌内注射）、低中高不同剂量（106、107、108）静脉注射多次治疗CBRH-7919接种的大鼠HCC模型，并设预治疗组、细胞因子加CBRH-7919细胞匀浆粗提物治疗组和生理盐水组作对照，用χ2检验比较其疗效和生存期。结果比较各组获得长期生存（生存大于100d）的大鼠数，细胞因子加CBRH-7919细胞匀浆粗提物治疗组、腹腔治疗组及高剂量静脉治疗组与生理盐水组比较无显著意义(P＞0.05)；中低剂量静脉治疗组、预治疗组及癌内注射组与生理盐水组比较有显著意义(P＜0.05、P＜0.001、P＜0.001和P＜0.01；预治疗组成癌率与其余各组相比有显著意义(P＜0.001)。结论中、低剂量负载癌抗原DC经静脉和癌内多次注射，对大鼠HCC有一定的疗效，并能有效地抵抗HCC细胞的再接种，提示负载癌抗原的DC治疗HCC具有潜在的临床应用前景。%Objectives　To study the feasibility of hepatocellular carcinoma immunotherapy with dendritic cells loaded with tumor antigens and to provide the fundation for its clinical application.Methods　Rat hepatocellular carcinoma models were treated with dendritic cells derived from rat bone marrow mononuclear cells pulsed with CBRH-7919 cellular homogenate extract by different way (administrated via tail vein,abdominal cavity,intratumoral) and with different cell quantity (10６，10７，10８) via vein.The tumorbearing rats in control groups were given NS and CBRH-7919 cellular homogenate extract and cytokines separately.The rats in another group which have been pre-treated with rat bone marrow derived syngeneic dendritic cells pulsed with CBRH-7919 cellular homogenate extract were challenged with 1×10６ CBRH-7919.The curative effect and the survival time of rats in each group was observed and compared with the control groups by χ2 test.Results　The survival rates of the rats in intravenous group (108) and in abdominal cavity group were not significantly different from control groups (compared with control group,P＞0.05 by χ2 test).The rats in intravenous groups (106,107) and in intratumoral group had significant prolonged survival (compared with control group,P＜0.05,P＜0.001 and P＜0.01 respectively,by χ2 test).Rats in pre-treated group did not develop tumors after being challenged with 1×10６　CBRH-7919 and had a longer survival(compared with control group,P＜0.001,by χ2 test).Conclusions　The immunotherapy of hepatocellular carcinoma rats with middle and low dose dendritic cells pulsed with cellular homogenate extract is effective relatively via vein and intratumor repeatedly.Dendritic cells loaded with tumor antigens can resist the challenge of reinoculability of hepatoma cells effectively.These results suggest that hepatocellular carcinoma immunotherapy with dendritic cells loaded with tumoral antigens has potential feasibility to be applied in clinical settings.