小剂量环磷酰胺通过抑制调节性T细胞提高小鼠肝癌阿霉素化疗效果  

Small -dose cyclophosphamide enhances chemotherapeutic effect of doxorubicin against mice hepatoma through inhibition of regulatory T cell

作　　者：沈顺利[1] 梁力建[1] 彭宝岗[1] 匡铭[1] 赖佳明[1] 黎东明[1] 

机构地区：[1]中山大学附属第一医院肝胆外科,广州510080

出　　处：《消化肿瘤杂志（电子版）》2009年第2期88-91,共4页Journal of Digestive Oncology(Electronic Version)

基　　金：国家自然科学基金(30872486);广东省自然科学基金(8151008901000096);广州市科技计划(2008B080703031);广东省医学科学技术研究基金WSTJJ20071116440102196306263231)

摘　　要：目的探讨小剂量环磷酰胺是否可以提高小鼠肝癌阿霉素化疗效果,并对其机制进行探讨。方法建立C57BL/6J小鼠皮下肝癌模型,待肿瘤长至100～150 mm^3体积大小时备用。10只小鼠随机分为2组,对照组不处理,观察组以2 mg/只的剂量腹腔注射环磷酰胺(CTX),4天后以流式细胞仪检测小鼠脾脏CD4^+、CD8^+T细胞和CD4^+CD25high Treg淋巴细胞比例。32只小鼠随机分为4组:①对照组(PBS);②环磷酰胺组(CTX);③阿霉素组(DOX);④环磷酰胺+阿霉素组(CTX+DOX);观察小鼠皮下肿瘤生长情况和小鼠的生存期。结果应用小剂量CTX后荷瘤小鼠脾脏CD4^+和CD8^+T淋巴细胞的浸润分别由17.62%、16.03%上升到24.54%和25.41%(P<0.05),而Treg则由用药前的11.92%降低为用药后的5.36%(P<0.05)。CTX组肿瘤生长和对照组相比无统计学差异(P>0.05)。DOX组、CTX+DOX组肿瘤生长均显著受到抑制,以CTX+DOX组更为显著(P<0.05)。DOX组生存期显著改善(P=0.027),联合应用CTX后生存期亦有显著提高(P=0.018)。结论小剂量CTX可以抑制小鼠脾脏Treg浸润,改善机体免疫微环境,并可以增强肝癌阿霉素化疗的效果。免疫化疗治疗晚期肝癌具有一定的应用前景。Objective To investigate whether small-dose cyclophosphamide(CTX) can enhance chemotherapeutic effect of doxorubicin against mice hepatoma and its possible mechanisms. Methods C57BL/6J subcutaneous hepatoma model was established and when the tumor volume reached to 100～150 mm^3,mice were divided into groups.Ten mice were randomly divided into two group.2 mg CTX was injected i.p and 4 days later flowcytometry was used to examine the percentage of CD4^+,CD8^+ lymphocytes and CD4^+CD25high regulatory T cell(Treg).Another 32 mice were randomly divided into 4 groups;(1 )Control group;(2)CTX group;(3) Doxorubicin group(DOX); (4)CTX + DOX group.Mice were examined for tumor growth and survival.Results After application of low-dose CTX,the infiltration of CD4^+ and CD8^+ T lymphocytes in mice spleen increased from 17.62%,16.03%to 24.54%and 25.41%(P<0.05) respectively,while Treg decreased from 11.92% to 5.36%(P<0.05).There was no significant difference in tumor growth between the control and CTX groups(P>0.05).Tumor growth of DOX group,CTX+DOX group were significantly inhibited, especially CTX +DOX group(P<0.05).Survival analysis showed that there was no significant difference in survival between the control and CTX groups.Survival of DOX group was significantly improved(P=0.027).Compared with DOX group,survival time of CTX+DOX group was significantly increased(P = 0.018).Conclusion Low-dose CTX can inhibit Treg infiltration in spleen and enhance chemotherapeutic effect of doxorubicin against mice hepatoma.Immuno-chemotherapy might represent a promising treatment modality for hepatocellular carcinoma.

关 键 词：原发性肝癌 调节性T细胞 免疫化疗 免疫微环境 

分 类 号：R735.7[医药卫生—肿瘤]