LHBs与C53蛋白在肝癌细胞系HepG2中的共定位及结合区域研究  

Study on the co-localization and interacting regions of LHBs and C53 protein in HepG2 cell line

作　　者：张锦前[1] 池频频[1] 任娜[1] 李贵[1] 吴淑玲[1] 温少芳[1] 王琦[1] 刘顺爱[1] 成军 

机构地区：[1]北京首都医科大学附属北京地坛医院,北京100015

出　　处：《国际病毒学杂志》2012年第4期156-160,共5页International Journal of Virology

基　　金：国家自然科学基金（No.30600524）

摘　　要：目的研究肝癌细胞系HepG2中乙型肝炎表面抗原大蛋白（hepatitis B virus large surface protein，LHBs）与C53蛋白是否存在共定位，查明LHBs与C53蛋白的结合区域。方法分别构建pEGFP-C1·LHBs和pDsRedl-N1·C53质粒，共同转染肝癌细胞系HepG2，激光共聚焦观察它们在细胞内的共定位。构建质粒pCDNA-3．1（-）myc／his-LHBs和pCMV-5a-C53，并将C53基因截短构建质粒pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2和pCMV．5a-C3，将pCDNA-3．1（-）myc／his．LHBs和pc-MV-5a-C53及各截短质粒共转染HepG2细胞，免疫共沉淀方法验证LHBs结合的C53区域。结果成功构建pEGFP-C1-LHBs、pDsRedl-N1-C53、pCDNA-3．1（-）myc／his-LHBs、pCMV-5a-C53、pCMV-5a-C53N、pCMV-5a-C1、pCMV-5a-C2和pCMV-5a-C3质粒；用激光共聚焦的方法证实了LHBs与C53在HepG2细胞内存在共定位；免疫共沉淀方法证实了在HepG2细胞内LHBs与C53的结合区域为其c1区。结论LHBs与C53在肝癌细胞系HepG2内存在明确的相互作用，但仍需做进一步的功能研究。Objective To identify the co-localization and interacting region of the hepatitis B virus large surface protein （LHBs） and C53 protein in HepG2 cell line. Methods The plasmids pEGFP-C1-LH- Bs, pDsRed1-N1-C53, pCDNA-3.1 （-） myc/his-LHBs, pCMV-Sa-C53, pCMV-Sa-C53N, pCMV-5a-C1, pCMV-Sa-C2 and pCMV-5a-C3 were constructed. The plasmids pEGFP-C1-LHBs and pDsRedl-N1-C53 were co-transfected into the HepG2 cells, and their co-localization was observed by laser scanning confocal microscope（LSCM）. The plasnaids pCDNA-3.1 （-） myc/his-LHBs and pCMV-5a-C53, pCMV-5a-C53N, pCMV-5a-C1, pCMV-5a-C2, pCMV-5a-C3 respectively were eo-transfected into the HepG2 cells, and the interacting region of LHBs and C53 was identified by eo-immunoprecipitation （CO-IP）. Results The plas- mids pEGFP-C1-LHBs, pDsRedl-N1-C53, pCDNA-3.1（-） myc/his-LHBs, pCMV-5a-C53, pCMV-5a-C53N, pCMV-5a-C1, pCMV-5a-C2 and pCMV-5a-C3 were constructed successfully. The co-localization of LHBs and C53 in HepG2 cells were identified, and the interacting region of LHBs and C53 was C1 of C53. Conclusion The interaction between LHBs and C53 was clear, and the further function research should be done.

关 键 词：乙型肝炎表面抗原大蛋白 C53 HEPG2 共定位 结合区域 

分 类 号：R512.62[医药卫生—内科学]