过表达Akt1细胞株的构建及鉴定  

Establishment and Identification of Over Expression of Akt1 Cell Strain

作　　者：潘姝花 郑婷婷[1] 郑旭升[1] 吴登伟[1] 陈培远[1] 许传莲[1] 

机构地区：[1]浙江理工大学生命科学学院,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2014年第4期462-466,共5页Journal of Zhejiang Sci-Tech University(Natural Sciences)

摘　　要：Akt1是细胞信号传导通路中的关键信号分子,具有促进细胞增殖、生长、迁移、侵袭,以及抑制细胞凋亡,抵抗化疗和放疗等重要作用。文章通过构建pLJM1-Akt1重组质粒,利用慢病毒侵染的方法将重组质粒转染至K562、Bel-7404细胞,用嘌呤霉素筛选得到Akt1稳定过表达的Bel-7404/Akt1、K562/Akt1细胞株,通过Western bloting分析细胞株中Akt1的表达情况。结果显示:Bel-7404/Akt1与K562/Akt1细胞中Akt1表达量明显高于野生型Bel-7404细胞与K562细胞,成功构建过表达Akt1的K562/Akt1、Bel-7404/Akt1稳转细胞株。Akt1过表达细胞株的成功构建为寻找和筛选高效、低毒、强特异性的Akt1抑制剂以及逆转细胞多药耐药(multidrug resistance,MDR)的研究提供了实验模型。Akt1 is a key signal molecule in cell signal transduction pathways,which promotes cell growth,proliferation,migration and invasion,inhibits cell apoptosis and resists chemotherapy and radiotherapy.This study establishes the pLJM1-Akt1 recombinant plasmids and transfects the recombinant plasmids into K562 and Bel-7404 cells with the method of virus infection.We obtained stable Bel-7404/Akt1 and K562/Akt1 over expressed by Akt1 through puromycin screening.Akt1expression in the cell strain was analyzed through Western bloting.The results show that the expression quantity of Akt1 in K562/Akt1 and Bel-7404/Akt1 cells is significantly more than that in wild K562 and Bel-7404 cells;stable and transfected cell strains of K562/Akt1 and Bel-7404/Akt1 of over expressed Akt1 are successfully established.The successful establishment of over expressed Akt1 cell strain provides experimental model for seeking and screening efficient,low-toxicity,strong-specificity Akt1 inhibitors and studying reverse multidrug resistance（MDR）.

关 键 词：AKT1 重组质粒 病毒侵染 

分 类 号：Q786[生物学—分子生物学]