人绒毛膜促性腺激素β亚基DNA疫苗的构建及其抗肿瘤作用的初步研究  

The Construction of Human Chorionic Gonadotropin β- Subunit Glycoprotein (hCGβ) DNA Vaccine and Its Preliminary Antitumor Study*

作　　者：陈云[1] 石树群[1] 杨颖[1] 彭景楩[1] 

机构地区：[1]中国科学院动物研究所

出　　处：《生物化学与生物物理进展》2003年第1期60-66,共7页Progress In Biochemistry and Biophysics

基　　金：国家高技术"86 3"计划资助项目 ( 2 0 0 1AA2 15 42 1);中国科学院知识创新工程重要方向项目 (KSCX.-SW- 2 0 1) .~~

摘　　要：人绒毛膜促性腺激素 β亚基 (hCGβ)除了在正常的妊娠滋养层细胞中分泌外 ,在许多肿瘤细胞也大量分泌 ,是防治hCGβ依赖型癌症的有效目标分子之一 .通过RT PCR方法克隆全长的hCGβ基因 ,成功构建了PCR3 1 hCGβDNA疫苗 ,其能够在HeLa细胞中高效表达hCGβ蛋白 ,表达的hCGβ蛋白主要存在于细胞内 .将2 0 μgPCR3 1 hCGβ质粒DNA通过普鲁卡因盐酸盐 (bupivacaine HCl)药物诱导后接种小鼠肌肉 ,小鼠能够吸收质粒DNA ,并表达编码的hCGβ蛋白抗原 ,表达的hCGβ被小鼠的免疫系统识别 ,同时激发hCGβ抗原特异性的体液免疫和细胞免疫反应 ,抗体滴度最高可超过 1∶80 0 0 ,并且这两种类型的免疫应答均能够在体外作用于HeLa细胞 ,诱导其发生细胞凋亡 ,表明PCR3 1 hCGβDNA疫苗激发的免疫反应在体外具有抗肿瘤作用 .The β- subunit of human chorionic gonadotropin (hCGβ) is secreted by trophoblastic cells during normal pregnancy. In addition, hCGβ is also synthesized by many tumor and cancer cells, and it has been one of target molecules for active immunotherapy to prevent and treat hCGβ dependent tumors and cancers. The full coding region of hCGβ was isolated by the RT- PCR method, and was inserted into PCR3 1 eukaryotic expression vector to construct the recombinant PCR3 1- hCGβ DNA vaccine successfully. Using HeLa cells transient expression system, the capability of hCGβ expression of PCR3 1- hCGβ was confirmed in vitro , and the expressed hCGβ protein was mainly with intracellular state. With 20 μg PCR3 1- hCGβ plasmid DNA through intramuscular inoculation after bupivacaine- HCl inducing, the immunized BALB/c mice could express hCGβ antigen to induce both intensive hCGβ- specific humoral immune responses and CTL responses, and the antibody titer reached high to 1∶8000. Moreover, both two types of immune responses elicited by PCR3 1- hCGβ could attack HeLa cells to induce apoptosis in vitro . The results indicated that the immune responses induced by PCR3 1- hCGβ DNA vaccine have antitumor effects in vitro , and would be helpful to detect the antitumor effects of PCR3 1- hCGβ DNA vaccine in vivo .

关 键 词：人绒毛膜促性腺激素β亚基 DNA疫苗 抗肿瘤作用 构建 表达 

分 类 号：R730.3[医药卫生—肿瘤]