preS1分子连接体转运siHBX抑制HepG2-HBX细胞株内HBX表达的实验研究  

prS1 molecule conjugate carrying siHBX inhibits expression of HBX in HepG2-HBX cells

作　　者：杨兵[1] 文阳安[1] 黄世峰[1] 周倩云[1] 张莉萍[1] 陈维贤[2] 

机构地区：[1]重庆医科大学附属第一医院检验科,重庆400016 [2]重庆医科大学附属第二医院检验科,重庆400010

出　　处：《第三军医大学学报》2011年第3期250-253,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30972582;30973378;81071621);重庆市科委项目(2009BB5276;2009BB5257;2010BB5390);重庆医科大学附属第一医院院内医学科学基金(YXJJ2009-12)~~

摘　　要：目的探讨应用preS1分子连接体作为siRNA转运载体对HepG2-HBX细胞株中稳定表达的乙型肝炎病毒X蛋白(HBX)的抑制效果及生物学效应。方法构建HepG2-HBX稳定表达细胞株;化学合成pres1分子连接体及HBX特异性siRNA序列(siHBX);pres1分子连接体分别与肝细胞系及siRNA进行结合能力试验,然后以preS1分子连接体作为转运载体转染siHBX入HepG2-HBX后经实时荧光RT-PCR及Western Blot检测siHBX对HBX基因的抑制效果。结果成功构建了稳定表达HBX基因的HepG2-HBX细胞株,preS1分子连接体能够与siRNA进行有效结合,同时可以携带siRNA进入肝细胞内高效抑制HepG2-HBX细胞株内HBX的表达。与空白组相比,阴性对照组抑制率为(11.94±6.3)%,而阳性对照组(脂质体)及实验组(preS1分子连接体)的抑制率分别为(85.24±2.6)%和(77.80±6.10)%,实验组与空白组及阴性对照组相比,差异均具有统计学意义(P<0.05)。结论 preS1分子连接体可作为一种新型siRNA转运载体,携带siHBX进入肝癌细胞内发挥RNA干扰机制,为乙型肝炎病毒相关性肝脏疾病的治疗提供了新的途径。Objective To study the inhibitory and biological effect of prS1 molecule conjugate carrying siHBX on expression of the HBX gene in HepG2-HBX cell line.Methods pres1 molecule conjugate and siHBX were chemically synthesized.Ability of the preS1 molecule conjugate to bind to HepG2-HBX and siRNA was tested.The siRNA was then transfected into the HepG2-HBX cell line by the preS1 molecule conjugate.Inhibiting effect of siHBX on expression of the HBX gene was detected by quantitative real-time RT-PCR and Western blotting,respectively.Results The HepG2-HBX cell line stably expressing the HBX gene was successfully constructed.The preS1 molecule conjugate could effectively bind to siHBX and inhibit expression of HBX in HepG2-HBX cells.The inhibition rate of pres1 molecule conjugate on expression of HBX was significantly lower in negative control groups than in postive and experimental group（11.94%±6.3% vs 85.24%±2.6% and 77.80%±6.1%,P0.05） Conclusion preS1 molecule conjugate carrying siHBX can be used as a novel siRNA carrier and plays a role in interference with RNA in liver cancer cells,thus providing a new treatment modality for HBV-related liver diseases.

关 键 词：preS1分子连接体 乙型肝炎病毒X蛋白 细胞穿透性多肽 靶向性配体 

分 类 号：R394.33[医药卫生—医学遗传学] R394.3[医药卫生—基础医学]