环磷酰胺联合照射后半相合淋巴细胞对小鼠肝癌移植瘤的抑制作用

Inhibitory effect of cyclophosphamide combined with irradiated haploidentical lymphocyte infusion on transplanted hepatocellular carcinoma in mice

目的:观察不同剂量环磷酰胺(cyclophosphamide,CTX)预处理联合5 Gy60Co照射的半相合淋巴细胞输注(hap-loidentical lymphocyte infusion,HLI)对小鼠肝癌移植瘤的抑制作用。方法:以皮下接种Hepa1-6肝癌细胞的BABL/c×C57BL杂交F1代雌性小鼠(表型为H-2b/d)为受鼠,以BALB/c×C3H杂交F1代雌性小鼠(表型为H-2d/k)为MHC半相合的供者,分PBS组、CTX 80 mg/kg+5 Gy照射HLI组、CTX 200 mg/kg+5 Gy照射HLI组、CTX 300 mg/kg+5 Gy照射HLI组、5 Gy照射HLI组,每组5只小鼠;观察各组小鼠瘤块生长和大小,检测受鼠体内的嵌合状态及移植物抗宿主病(graft-versus host disease,GVHD)的发病情况。结果:80、200 mg CTX联合HLI组小鼠瘤体积小于PBS组[(1.25±0.24)、(1.38±0.31)vs(2.03±0.24)cm3,P<0.01],小鼠生存时间显著长于PBS组[48 d(39 d,55 d)、40 d(35 d,48 d)vs 35 d(18 d,39 d),P<0.05];80mg CTX联合HLI组的抑瘤作用强于单纯HLI组[(1.25±0.24)vs(1.76±0.40)cm3,P<0.05];300 mg CTX联合HLI组和单纯HLI组无明显抑瘤作用。各治疗组小鼠均未出现GVHD。80、200 mg CTX联合HLI组小鼠嵌合度低于300 mg CTX联合HLI组,且消失时间明显早于后者。结论:低剂量CTX联合输注经照射的半相合供者淋巴细胞可获得较好的抗小鼠肝癌移植瘤的作用,CTX剂量增加后抗肿瘤作用并未增强。

Objective： To analyze the effect of irradiated haploidentical lymphocyte infusion（HLI） following different doses of cyclophosphamide（CTX） on hepatocellular carcinoma Hepal-6 cells.Methods： Female（BABL/c×C57BL）→F1（CB6F1 H-2b/d） mice were subcutaneously inoculated with Hepa1-6 cells to develop a solid tumor model that was taken as recipients.The irradiated haploidentical lymphocytes were from the female（BALB/c×C3H）→F1（CC3HF1 H-2d/k）.All the mice were divided to five groups： group PBS,group CTX 80 mg/kg＋SP/irr（spleen lymphocyte/5Gy irradiated）,group CTX 200 mg/kg＋SP/irr,group CTX 300 mg/kg＋SP/irr and group SP/irr.The tumor size was measured every 2-3 days.The chimerism was measured after HLI,and the graft-versus host disease（GVHD） was assessed.Results： The tumor was significantly suppressed in group CTX 80 mg/kg＋SP/irr and group CTX 200 mg/kg＋SP/irr compared with group PBS（[1.25±0.24] cm3,[1.38±0.31] cm3 vs [2.03±0.24] cm3,P0.01）,so was the survival time（48 d [39 d,55 d],40 d [35 d,48 d] vs 35 d [18 d,39 d],P0.05）,of which,group CTX 80 mg/kg＋SP/irr got significantly different antitumor effect from the group SP/irr（[1.25±0.24] cm3 vs [1.76±0.40] cm3,P0.05）.GVHD was not found in any groups.The chimerism of group 80 mg and 200 mg CTX combining with HLI was lower than group CTX 300 mg combining with HLI and the disappearence time was shorter.Conclusion： Transplantation of irradiated haploidentical lymphocytes following low does of CTX can induce antitumor effect on hepatocellular carcinoma of mice.Higher doses of CTX may not increase antitumor effect.