人血型B抗原模拟多肽、Mip3β双表达重组质粒体外转染人黑色素瘤细胞B16的抗肿瘤效应  

Antitumor effect of recombinant plasmid expressing peptide mimic of blood type B antigen and Mip3β after transfecting human melanoma cell line B16

作　　者：罗敏[1] 周媛[1] 岑东芝[1] 张积仁[1] 

机构地区：[1]南方医科大学珠江医院肿瘤中心,广州市510282

出　　处：《实用医学杂志》2010年第18期3320-3322,共3页The Journal of Practical Medicine

基　　金：卫生部科技专项基金(编号:W2009BX015)

摘　　要：目的:将人血型B抗原模拟多肽、Mip3β双表达重组质粒于人黑色素细胞B16转染,检测其对抗肿瘤效应的影响。方法:选用人黑色素瘤细胞株B16,设4个组:A组,转染P1/Fas-Mip3β-pIRES组;B组,转染P1/Fas-pIRES组;C组,转染Mip3β-pIRES组;D组,转染pIRES组。转染方法采用Invitrogen(美国)公司的LipofectamineTM2000脂质体转染的方法(6孔板,8×105孔,2mL体系)。分别流式细胞仪检测细胞凋亡率,进行体外补体介导的细胞杀伤(CDC)实验和抗体依赖细胞介导的细胞毒作用(ADCC)实验,获得细胞活力并进行统计学分析。结果:重组质粒成功转染人黑色素瘤细胞株B16:(1)流式细胞仪检测双表达组组细胞凋亡率87.6%,明显较其他组别高。(2)CDC试验发现最小均值的组合为20g/L组B肽-MIP-pIRES。(3)ADCC试验发现最小均值的组合为40∶1组B肽-MIP-pIRES。结论:人血型B抗原模拟多肽、Mip3β双表达重组质粒能在人黑色素细胞B16转染,诱导肿瘤细胞凋亡,并通过CDC及ADCC效应增强免疫反应。Objective To explore the antitumor effect of recombinant plasmid expressing peptide mimic of blood type B antigen and Mip3β after transfecting human melanoma cell line B16 in vitro. Methods With lipofectamine TM2000, human uman melanoma cell line B16 was transfected by P1/Fas-Mip3β-pIRES （group A）, P1/Fas-pIRES （group B）,Mip3β-pIRES （group C）, and pIRES （group D）, respectively. Apoptotic rate was detected by flow cytometry. Complement-mediated cytotoxicity （CDC） assay and antibody-dependent cell-mediated cytotoxicity （ADCC） assay were conducted to measure cell viability. The date was statistically analyzed by SPSS 10. Results The recombinant plasmid was successfully transfected melanoma cell line B16. The apoptotic rate was greater in group A （87.6%） than in the other groups. CDC test showed the minimum mean value was 20 μg/uL in the group with B peptide-MIP-pIRES; and ADCC test showed the minimum mean value was 40 ∶ 1 in the group with B peptide-MIP-pIRES. Conclusions The recombinant plasmid expressing peptide mimic of blood type B antigen and Mip3β can successfully transfect human melanoma cell line B16, induce apoptosis of the tumor cells, and enhance immune reaction by complement-mediated cytotoxicity and antibody-dependent cell-mediated cytotoxicity.

关 键 词：黑色素瘤 血型抗原 Mip3β 重组质粒 

分 类 号：R739.5[医药卫生—肿瘤]