重组质粒pVAX1-HER2-Flt3L构建及抗肿瘤效果  

Construction and anti-tumor effect of recombinant plasmid pVAX1-HER2-Flt3L

作　　者：杨克科[1] 石磊[2] 范永鹏 陈景华[1] 

机构地区：[1]黑龙江中医药大学佳木斯学院,黑龙江佳木斯154007 [2]佳木斯大学附属第一医院,黑龙江佳木斯154000 [3]中国农业科学院农产品加工研究所主食加工技术研究所,黑龙江哈尔滨151900

出　　处：《生物技术》2017年第6期517-521,共5页Biotechnology

基　　金：黑龙江省自然科学基金项目("马钱苷和莫诺苷抗恶性黑素瘤的作用机制研究";No.D201275)

摘　　要：[目的]将具有免疫增效功能的Flt3L分子与HER2抗原融合表达,评价Flt3L是否能够提高抑瘤效果。[方法]将人的HER2编码区基因和小鼠Flt3L编码区基因用Furin/2A进行连接,全基因合成后,克隆至pVAX1载体,从而构建重组质粒pVAX1-HER2-Flt3L,同时构建对照质粒pVAX1-HER2和pVAX1-Flt3L。将上述质粒分别注射正常小鼠和稳定表达HER2抗原的荷瘤小鼠,评价其免疫原性和抑瘤效果。[结果]将各组重组质粒接种于Bal B/c小鼠后,抗体检测结果显示:重组融合质粒pVAX1-HER2-Flt3L组诱导产生的平均抗体滴度为2060,而单纯抗原质粒pVAX1-HER2组诱导产生的平均抗体滴度为1720;ELISPOT检测结果显示:重组融合质粒pVAX1-HER2-Flt3L组诱导产生的平均斑点数为133,而单纯抗原质粒pVAX1-HER2组诱导产生的平均抗体滴度为110;与此一致的是,在荷瘤小鼠中,该融合质粒也显示出更加显著的抑制肿瘤生长的效果,达到47%,而单纯抗原质粒仅为32%。[结论]Flt3L具有一定的免疫增效能力,能够显著提高表达HER2抗原重组质粒的抑瘤效果,深入研究其免疫保护机制具有潜在的临床意义。[Objective]The Flt3L molecule was fused with the HER2 antigen to evaluate whether the Flt3L molecule could improve the anti-tumor effect.[Methods]The HER2 encoding gene and mouse Flt3L gene encoding was linked through Furin/2 A.After gene synthesis,it was cloned into vector pVAX1 to construct recombinant plasmid pVAX1-HER2-Flt3L,and control plasmid pVAX1-HER2 and pVAX1-Flt3L were also constructed.The plasmids were injected into normal mice and tumor bearing mice stably expressing HER2 antigen,and their immunogenicity and antitumor effect were evaluated.[Results]After recombinant plasmids were injected into Bal B/c mice,antibody test results showed that the average antibody titer induced by fusion plasmid pVAX1-HER2-Flt3L group was 2060,compared with alone antigen plasmid pVAX1-HER2 group was 1720.The average number of spots induced by the recombinant fusion plasmid group was 133,while the average number of spots induced by the alone antigen plasmid group was 110; consistent with this,in tumor bearing mice,the fusion plasmid group also showed a more significant effect on inhibition tumor growth,up to 47%,while only 32% of the alone antigen plasmid group.[Conclusion]Flt3L has a certain immune synergistic effect,which can significantly improve the anti-tumor effect of recombinant plasmid expressing HER2 antigen,and further study of its immune protection mechanism has potential clinical significance.

关 键 词：人表皮生长因子受体2 FMS相关的酪氨酸激酶3配体 肿瘤 结肠癌 

分 类 号：R73-362[医药卫生—肿瘤]