负载肝癌细胞裂解物的树突状细胞Transgelin表达水平与其功能的关系

摘要

目的 探讨负载肝癌细胞裂解物的树突状细胞(DC)Transgelin表达与其表型和功能的关系.方法 正常人来源的DC分成3组,分别为负载高转移潜能肝癌细胞MHCC97H、低转移潜能肝癌细胞MHCC97L的裂解物组和无负载DC(对照)组.用共聚焦显微镜和扫描电子显微镜观察DC形态;流式细胞仪检测DC表型,混合淋巴细胞反应及其上清液中白细胞介素(IL)-12含量测定以反映DC功能;Western blot测定DC中Transgelin的表达.结果 3组DC的形态未有明显不同,负载低转移潜能肝细胞癌组DC的CD80、CD83、CD86、混合淋巴细胞反应和IL-12含量明显高于对照组(P<0.01);负载高转移潜能肝细胞癌组DC的CD80、CD86、Transgelin表达明显高于对照组(P<0.05);负载高转移潜能肝细胞癌组DC的CD80、CD83、CD86和IL-12含量明显低于低转移组(P<0.05),而Transgelin表达则高于低转移组.结论 负载肝细胞癌裂解物的DC中Transgelin表达与其表型和功能有关.%Objective To explore the relationships between the expression of transgelin in dendritic cells (DCs) pulsed with hepatocellular carcinoma lysates and the functions of the DCs. Methods DCs derived from healthy human white blood cells were divided into 3 groups: one was pulsed with high meta- static potential hepatocellular carcinoma cell line (MHCC97H) lysates, one with lysates of a low metastatic potential cell line (MHCC97L), and one un-pulsed DCs served as the control. The morphology of the DCs was observed by confocal microscopy and scanning electron microscopy. The phenotypes of the DCs were detected by flowcytometric analysis. The mixed leucocyte reaction (MLR) test and IL-12 secretion of DCs in the supernatants of MLR were employed to determine the functions of the DCs;the expression of transgelin was detected by Western blot. Results There were no morphological changes in the different DCs, but the levels of HLA-DR, CD80, CD83, CD86, MLR and IL-12 and transgefin were significantly higher in the two pulsed groups than those in the control group (P < 0.01). In MHCC97H pulsed DCs, their CD80, CD83, CD86, and the expression of transgelin were also higher than those in the control group (P < 0.05). The expression of transgelin was significantly higher in the MHCC97H pulsed group than in the MHCC97L loaded group, but CDS0, CD83, CD86 and the level of IL-12 were all lower in the MHCC97H loaded DC group in comparison with those in the MHCC97 pulsed group (P < 0.05). Conclusion The expression of transgelin in DCs pulsed with HCC lysates is related to the functions of the DCs.