重组人粒细胞-巨噬细胞集落刺激因子溶瘤2型单纯疱疹病毒注射液(Vero细胞)的药效学分析  

Pharmacodynamic of recombinant human granulocyte-macrophage colony stimulating factor oncolytic herpes simplex virus type 2 (Vero cells) injection

作　　者：金静 汪洋[1] 胡翰 毛泽勇 石晓太 方志正 刘滨磊[1] JIN Jing;WANG Yang;HU Han;MAO Ze-yong;SHI Xiao-tai;FANG Zhi-zheng;LIU Bin-lei(Hubei University of Technology, Wuhan 430000, Hubei Province, Chin)

机构地区：[1]湖北工业大学,湖北武汉430000 [2]武汉滨会生物科技股份有限公司,湖北武汉430000

出　　处：《中国生物制品学杂志》2018年第5期502-506,共5页Chinese Journal of Biologicals

摘　　要：目的分析重组人粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)溶瘤2型单纯疱疹病毒(oncolytic herpes simplex virus type 2,OH2)的体内、外药效。方法按不同感染复数(MOI)=0.002、0.01、0.05、0.25、1.25分别感染人结肠癌细胞(Lo Vo)和人喉癌细胞(Hep2),MTT法检测OH2在体外的溶瘤作用。经BALB/c-nu正常小鼠右侧肋腹皮下分别注射Lo Vo细胞和Hep2细胞,当肿瘤平均体积>100 mm^3时,试验组于肿瘤内分别注射高、中、低剂量的OH2(1×10~7、1×10~6、1×10~5 CCID_(50)/m L),同时设阳性对照组(5 mg/m L的5-FU)及阴性对照组(DMEM/F12培养基),于第0、3和6天进行注射。首次注射后第0、3、6、9、12、15、18天测量肿瘤长径(a)和宽径(b),计算肿瘤体积(tumor volume,TV)及相对肿瘤增殖率。结果 OH2在较小浓度下(MOI分别为0.168和0.063)即可较好地杀伤Lo Vo和Hep2细胞。与阴性对照组比较,两种肿瘤模型的OH2高、中、低剂量组及阳性对照组均有显著抑瘤效果(P<0.05);Lo Vo肿瘤模型中,OH2高剂量组的抑瘤效果明显优于中、低剂量组(P<0.05);Hep2肿瘤模型中,OH2高、中、低剂量组的抑瘤效果差异无统计学意义(P>0.05)。结论 OH2在体外和体内实验中均显示较好的溶瘤作用,具有新生物药开发前景,为临床前研究提供了实验依据。Objective To analyze the in vivo and in vitro pharmacodynamics of recombinant human granulocyte-macrophage colony stimulating factor（GM-CSF）onxolytic herpes simplex virus type 2（OH2）. Methods Human colon cancer Lo Vo cells and human laryngeal carcinoma Hep2 cells were infected with OH2 at various MOIs（0. 002,0. 01,0. 05,0. 25 and 1. 25）. The oncolytic effect of OH2 in vitro was determined by MTT assay. BALB/c-nu mice were injected s. c. with Lo Vo and Hep2 cells respectively to establish the tumor models. After the sizes of tumors were more than100 mm-3,the OH2 were injected into the tumors at high（1 × 10^-7 CCID（50）/m L）,moderate（1 × 10-6 CCID（50）/m L）and low（1 × 10^-5 CCID（50）/m L）respectively,each on days 0,3 and 6,using 5 mg/m L 5-FU as positive control and DMEM/F12 medium as negative control. The longitudinal（a）and transverse（b）diameters of tumors were measured on days 0,3,6,9,12,15 and 18 after the first injection,based on which the tumor volume（TV） and relative tumor proliferation rate were calculated. Results OH2,even at a low concentration,killed Lo Vo and Hep2 cells（at MOIs of 0. 168 and0. 063） effectively. Compared with that in negative control group,obvious tumor-inhibiting effects were observed in the high,moderate and low dose OH2 as well as positive control groups in the two tumor models（P〈0. 05）. In the mice injected with Lo Vo cells,the tumor-inhibiting effect in high dose OH2 group was superior to those in moderate and low dose OH2 groups（P〈0. 05）. However,in the mice injected with Hep2 cells,the tumor-inhibiting effects of OH2 at three dosages showed no significant difference（P〉0. 05）. Conclusion OH2 showed good oncolytic activity in both in vitro and in vivo tests,which was of prospect of being developed as a novel biopharmaceuticals. It provided an experimental basis for preclinical study of OH2.

关 键 词：重组人粒细胞-巨噬细胞集落刺激因子 溶瘤2型单纯疱疹病毒 人结肠癌细胞 人喉癌细胞 药效学 

分 类 号：R373.13[医药卫生—病原生物学] R181.8[医药卫生—基础医学]