β-葡萄糖神经酰胺与HBsAg-DC瘤苗协同治疗肝癌的实验研究

Experimental study of β-GC combined with HBsAg gene-modified dendritic cell-based tumor vaccine to hepatocellular carcinoma by different immune ways

目的探讨β-葡萄糖神经酰胺(β-GC)联合乙型肝炎表面抗原(HBsAg)基因修饰树突状细胞(dendritic cell,DC)瘤苗治疗肝癌的作用。方法以重组腺病毒为载体构建HBsAg-DC瘤苗。将C57BL/6J小鼠随机分为6组(A-F,6只/组),其中A、B、C组接种PBS液2次,D、E、F组接种HBsAg-DC瘤苗2次。皮下注入HepG222.1.5肝癌细胞当天始,B、E组接受β-GC(1.5μg)腹腔注射,C、F组接受β-GC(15μg)灌胃给药,A、D组为安慰剂对照,比较不同组间移植瘤生长情况。结果 B、C组移植瘤生长较之A组明显受抑(P<0.05),E、F组移植瘤生长较之D组明显受抑(P<0.05)。A-F组移植瘤大小(mm3)分别为364.2±3.06,236.5±8.96,251.0±5.76,75.0±5.9,35.3±4.46,38.5±5.47。经腹腔注射给药与灌胃给药相比在抗瘤作用方面差异无统计学意义。结论β-GC经腹腔或灌胃给药均具有提高HBsAg-DC瘤苗的免疫治疗作用,其协同抗肿瘤效应的机制可能通过激活NKT细胞。

Objective To explore the efficacy of β-GC combined with HBsAg gene-modified DC vaccine on animal model with HCC using different immunization routes. Methods Recombinant adenovirus vector-mediated HBsAg gene-modified DC was used to construct HBsAg-DC which was infused into C57BL/6J mice bearing HBsAg-related HCC by subcutaneous injection. Mice in group A, B and C were vaccinated with PBS, group D, E and F were vaccinated with pAd-HBsAg-DC twice before HepG2 22.1.5 inoculation as described above. Starting at the day of inoculation, mice were treated with daily intra-peritoneal. （ip, 1.5 μ g per mouse） β-GC injection in group B and E or daily oral β-GC doses（po, 15 μg per mouse） in group C and F. Group A and D receiving vehicle （po） were set as control for 13 -GC treatment.The effectiveness of the immune therapy on tumor growth was compared among different groups. Results Tumor size measured 36 days after inoculation was （364.2 ± 3.06）mm3 in group A, （236.5 ± 8.96）mm3 in group B, （251.0 ± 5.76）mm3 in group C, （75.0 ± 5.9）mm3 in group D, （35.3 ± 4.46）mm3 in group E, and（38.5 ± 5.47）mm3 in group F. The application of β-GC demonstrated anti-tumor activity by itself （group B and C）, and also enhanced the tumor preventive effect of pAd-HBsAg-DC （group E and F）. No difference in tumor growth was observed between ip and po application of β-GC. Conclusion β-GC may serve as an immune enhancer to augment the anti-tumor immunity triggered by HBsAg gene-modified DC vaccine. It＇s antitumor effect may be related to activation of NKT cells.