电脉冲刺激法提高小鼠miR-122表达的研究  

Improving the Expression of miR- 122 Following Injected miR- 122 Plasmid DNA into Mice by Electric Pulse Method

作　　者：吴艳玲[1] 夏丽洁[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046

出　　处：《生物技术》2014年第4期71-75,共5页Biotechnology

基　　金：新疆维吾尔自治区高技术研究发展计划项目(编号:201110101)资助

摘　　要：目的:电脉冲能够改变细胞膜的通透性,促进外源DNA进入细胞内。通过基因导入仪电脉冲刺激已注入质粒pcDNA3.0-miR-122的小鼠肌肉细胞,检测对小鼠miR-122表达的影响。方法:将扩增得到的pre-miR-122克隆到真核表达载体pcDNA3.0上,构建pcDNA3.0-miR-122表达质粒。利用基因导入仪分别将生理盐水、pcDNA3.0、pcDNA3.0-miR-122质粒DNA导入小鼠腿部肌肉,然后进行电脉冲刺激。qRT-PCR测定各组小鼠血清及肝脏中的miR-122水平,同时检测肝癌组织和经Cecropin XJ处理的肝癌组织中的miR-122水平。结果:基因导入仪电脉冲刺激已注入pcDNA3.0-miR-122的腿部肌肉组织后,小鼠血清和肝脏中miR-122的表达分别提高了23和11倍,经Cecropin XJ处理的小鼠肝癌组织中miR-122的表达比对照高1.6倍。结论:电脉冲刺激能够显著提高小鼠体内miR-122的表达,为探讨miR-122表达对肝癌生长影响的调控作用奠定基础。Objective：To improve the expression of miR - 122 following injected miR - 122 plasmid DNA into mice by electric pulse meth- od. Method：The 244 bp of pre - miR - 122 gene fragment was cloned into pcDNA3.0,and constructed a pcDNA3.0 - miR - 122 expres- sion vector. The saline,pcDNA3.0 and pcDNA3.0 - miR - 122 plasmid were injected intramuscularly into mice by electric pulse meth- od. qRT - PCR was used to detect the change of miR - 122 expression levels in serum and liver of mice. Meanwhile,the expression levels of miR- 122 were also detected in hepatic carcinoma of the control and Cecropin XJ treatment groups ,respectively. Result：The expression levels of miR - 122 significantly increased after injected peDNA3.0 - miR - 122 in serum and liver of mice and in hepatic carcinoma of Cecropin XJ treatment. Condusion：The plasmid peDNA3.0 - miR - 122 was injected into mice by electric pulse method, and the expres- sion levels of miR - 122 in mice could significantly increased. It would help to elucidate the regulatory relationship between miR - 122 ex- pression and hepatic carcinoma growth

关 键 词：小鼠 pcDNA3.0-miR-122 电脉冲法 miR-122表达 肝癌 

分 类 号：Q782[生物学—分子生物学] Q812