DC荷载α-GalCer对小鼠HepS肝癌的抑制作用

Inhibition of DC load α-Gal Cer on HepS Hepatocellular Carcinoma in Mice

目的:研究树突状细胞(DC)荷载α-半乳糖神经酰胺(α-Gal Cer)作为自然杀伤T细胞(NKT)刺激物对原发性肝癌Heps荷瘤小鼠的抑制作用。方法:建立小鼠Hep S肝癌移植瘤模型,随机分为3组,对照组(NS对照组)、树突状细胞组(DC组)、DC荷载α-半乳糖神经酰胺组(DC+α-Gal Cer组),每组10只。分别给予尾静脉注射DC、荷载α-Gal Cer的DC4×106/m L,对照组给予同体积的NS,隔天给药1次,共3次。各组于干预后2周取眼球血,用流式细胞术(FCM)检测小鼠外周血CD4+T细胞及CD8+T细胞含量变化。取肿瘤组织,称取各瘤体重量,计算抑瘤率。免疫组化染色比较各组瘤体细胞内Bax凋亡蛋白;Westem blot法检测各组移植瘤组织内Bax的表达。结果:DC荷载α-Gal Cer组CD4+T细胞及CD8+T细胞数较DC组及对照组明显升高(P<0.05),DC组较对照组有上升趋势,但差异无统计学意义(P>0.05)。DC荷载α-Gal Cer组平均瘤体重量较对照组及DC组均显著减轻(P<0.05),DC组与对照组平均瘤重无明显变化(P>0.05)。与NS组及DC组相比,DC荷载α-Gal Cer组移植瘤内Bax阳性细胞数量及Western blot表达量明显升高,差异有统计学意义(P<0.05);DC组与NS组之间无明显差异(P>0.05)。结论:DC荷载α-Gal Cer可活化NKT细胞,激活外周血抗肿瘤免疫细胞杀伤肿瘤,抑制小鼠肝癌移植瘤的生长。

Objective： To investigate the inhibitory effects of DC load α-galactosylceramide as the stimulus of natural killer T cells（NKT） on liver cancer of mice. Methods： Mouse model of the solid tumor of liver cancer were established. Then the mice were randomly divided into three groups（n=10 each）： Normal saline control group, dendritic cells（DC） group and DC load α-Galcer group. For the latter two groups, 4 ×106/m L dentritic cells, DC cells loaded with α-Galcer were respectively injected into the mice through the tail vein,while the mice in the NS control group were administrated with the same volume of NS. The administration was carried out every other day, totally 3 times. Peripheral blood and spleen was detached 2 weeks after injection in each group. The number of CD4＋, CD8＋T cells in peripheral blood of mice was counted by flow cytometry（FCM）, The expression of Bax in transplanted tumor was detected by immunohistochemistry and Western blot. Results： The number of CD4＋and CD8＋T cells was higher in the DC load α-Galcer group than that of DC group and control group（P0.05）, while it was higher in DC group than that of the control group, but no obvious difference（P0.05）. The average tumor weight of DC load α-Galcer group was significantly decreased than that of DC group and the control group（P0.05）. There was no significant change in the average tumor weight between the DC group and control group（P0.05）. The number of Bax-positive cells in DC load α-Gal Cer group was significantly decreased compared with that of DC group and control group（P0.05）by Western blot detection, while no significant change was detectable in the number of Bax-positive cells between DC group and control group. Conclusions： DC load α-Galcer can activate NKT cells, and also can activate the peripheral blood anti-tumor immune cells to inhibit the growth of Heps tumor in mice.