抑制Treg联合瘤内转染Ad.GM-CSF增强阿霉素治疗肝癌效果的实验研究  

Combination of Treg inhibition and intratumoral transfection of Ad.GM-CSF enhances the chemotherapeutic effect of doxorubicin against hepatocellular carcinoma

作　　者：赖佳明[1] 沈顺利[1] 彭宝岗[1] 李薇[1] 杨广鑫[2] 冯允[2] 梁力建[1] 

机构地区：[1]中山大学附属第一医院肝胆外科 [2]中山大学中山医学院,广东广州510080

出　　处：《中国病理生理杂志》2010年第A10期1879-1884,共6页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.8151008901000161);教育部博士点基金资助项目(No.20070558233);国家大学生创新训练计划资助项目(No.091055846)

摘　　要：目的:探讨抑制调节性T细胞(Treg)联合瘤内转染Ad.GM-CSF增强阿霉素治疗肝癌效果的可行性,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100-150mm3时随机分为6组,每组12只:(1)对照组(PBS);(2)环磷酰胺组(CTX);(3)阿霉素组(DOX);(4)Ad.GM-CSF组;(5)CTX+DOX组;(6)CTX+DOX+Ad.GM-CSF组。末次治疗后第5d每组处死其中4只小鼠,取脾脏测定CTL活性;取肿瘤组织行病理检查;每组另外的8只小鼠用于观察皮下肿瘤生长和小鼠生存期。结果:CTX可以显著增强DOX对肿瘤生长的抑制作用(P<0.05),延长小鼠生存期[(62.13±4.21)dvs(79.88±9.00)d,P<0.05],而联合应用Ad.GM-CSF可显著增强该效果[(79.88±9.00)dvs(106.13±5.23)d,P<0.01]。同其它组相比,CTX+DOX+Ad.GM-CSF组小鼠瘤体内可见大量肿瘤细胞坏死,CD8+T细胞浸润显著增加,而小鼠脾脏CTL杀瘤活性显著增强(P<0.05)。结论:以阿霉素为治疗基础的化疗联合抑制Treg并瘤内转染Ad.GM-CSF的免疫治疗具有协同抗肝癌作用,免疫化疗治疗晚期肝癌具有一定的应用前景。AIM:To explore the possibility that combination of regulatory T-cell(Treg)inhibition and intratumoral transfection of Ad.GM-CSF enhances the chemotherapeutic effect of doxorubicin against hepatocellular carcinoma(HCC).METHODS:C57BL/6J subcutaneous HCC model was established.The mice were randomly divided into 6 groups with 12 mice in each group when the tumor volume reached to 100-150 mm3:control group,cyclophosphamide(CTX)group,doxorubicin(DOX)group,Ad.GM-CSF group,CTX+DOX group and CTX+DOX+Ad.GM-CSF group.Four mice in each group were sacrificed for spleen cytotoxic T-cell(CTL)activity assay and tumor tissue histological examination 5 d after the final therapy.Eight mice in each group were examined for tumor growth and survival.RESULTS:CTX enhanced the chemotherapeutic effect of doxorubicin against HCC by inhibiting the tumor growth(P<0.05)and prolonging the mice survival(62.13 d±4.21 d vs 79.88 d±9.00 d,P<0.05),which was significantly strengthened by the combined use of Ad.GM-CSF(79.88 d±9.00 d vs 106.13 d±5.23 d,P<0.01).Compared with other groups,more tumor necrosis in tumor specimens and more infiltration of CD8+ T-lymphocytes in CTX+DOX+Ad.GM-CSF group were observed.Furthermore,the spleen cytotoxic T-cell(CTL)activity was significantly improved(P<0.05).CONCLUSION:Doxorubicin-based chemotherapy and immunotherapy by the method of Treg inhibition combined with intratumoral transfection of Ad.GM-CSF have synergistic effect against HCC.

关 键 词：癌 肝细胞 T淋巴细胞 调节性 免疫化疗 多柔比星 

分 类 号：R735.7[医药卫生—肿瘤]