叶酸受体在血液肿瘤细胞株的表达及叶酸-聚乙烯亚胺共聚物的制备  

Expressions of folate receptor in hematology malignant cell lines and synthesis of folate-polyethylenimine

作　　者：韩新爱[1] 曾慧兰[1] 古晨[2] 蒋建伟[3] 钟启[1] 朱海扬[4] 

机构地区：[1]暨南大学附属第一医院血液科,广东广州510630 [2]中山大学医学院临床医学系,广东广州510089 [3]暨南大学医学院生化教研室,广东广州510632 [4]嘉应学院医学院药理教研室,广东梅州514031

出　　处：《暨南大学学报（自然科学与医学版）》2009年第6期658-663,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省自然科学基金博士启动基金项目(06300580);广州市番禺区科技计划(2008-z-64-1)

摘　　要：目的：研究叶酸受体（folatereceptor，FR）在血液肿瘤细胞表达的特点，合成叶酸-聚乙烯亚胺（FA—PEI）聚合物，探讨其作为靶向性基因输送载体的可行性。方法：用实时荧光定量PCR（realtimePCR）法检测α、β、γ3种FR在血液肿瘤细胞K562、K562／A02、U266细胞株上的表达；利用叶酸（FA）活性酯在微碱性条件下与聚乙烯亚胺（polyethylenimine，PEI）的支链氨基反应，合成FA-PEI共聚物；用葡聚糖凝胶柱G-25分离、纯化；用紫外分光光度计波长扫描法及氢核磁共振谱（1HNMR）法验证交联是否成功。结果：α、β、γ3种FR在K562、K562／A02、U2663种细胞株上均表达，其中，α-FR的表达占优势，较β-FR和γ-FR表达量高；紫外分光光度计扫描图谱在365nm处出现叶酸的特征性吸收峰；核磁共振（1HNMR）示：在2．5～3．2处出现PEI亚甲基质子的特征性化学位移，在6．5～9．0处出现叶酸芳香质子的特征性氢信号。结论：FR在K562、K562／A02、U2663种细胞株上均有较高表达；FA—PEI偶联成功，为其作为血液肿瘤治疗中1种潜在的靶向性基因输送载体提供了依据。Aim：Examining the expression patterns folate receptor in hematology malignant cell lines and synthesis a new potential targeted gene carrier vector folate-polyethylenimine. Methods： （1）Real-time fluorescence quantitative PCR was used to detect the expression patterns of folate receptor in three kinds of hematology tumor cell lines. （2） The activated folate （N-hydroxysuccinimide ester of folie acid） was eonjuated to the surface of PEI via the branched amino groups. Then the FA-PEI eopolymer was purified with Sephadex G-25 column and completely separated from unreacted folate. Folate-polyethylenimine was determined by ultraviolet （UV） spectrophotometric analysis and hydrogen nuclear magnetic resonance gy tumor cell lines K562, K562/A02, U266 and the expression levels of α-FR was significantly higher than those ofβ-FR and γ-FR. Thus the expression of α-FR was dominant. （2） Ultraviolet （UV） spectrophotometric analysis found that the isolated sample displays the characteristic absorption peak of folate in 365 nm. （3）1HNMR test showed that the characteristic chemical shifts of methylene proton was in 2. 5 ~ 3.2 and the characteristic aromatic hydrogen proton signal of folic acid was in 6. 5 - 9.0. Conclusions： （1） the expression levels of α-FR was higher than those ofβ-FR and γ-FR in the three hematology tumor cell lines. （2）The successful coupling of FA-PEI can be used as a potential targeted gene delivery vector and providing evidence in the treatment of hematology tumor.

关 键 词：叶酸受体 叶酸-聚乙烯亚胺共聚物 K562细胞 K562/A02细胞 U266细胞 

分 类 号：R733.3[医药卫生—肿瘤] R979.1[医药卫生—临床医学]