人ERK2基因对人视网膜母细胞瘤WERI-Rb-1细胞生长的影响  

Effect of Human ERK2 Gene on Growth of Human Retinoblastoma WERI-Rb-1 Cells

作　　者：韩笑 刘莹 梁迎春[2] 闫志风[3] 徐小洁[2] 梁九思 洪甜[2] 尤文叶 陈思禹[3] 黄俊[2] 叶棋浓[2] 刘丹 HAN Xiao;LIU Ying;LIANG Ying-Chun;YAN Zhi-Feng;XU Xiao-Jie;LIANG Jiu-Si;HONG Tian;YOU Wen-Ye;CHEN Si-Yu;HUANG Jun;YE Qi-Nong;LIU Dan(The First Affiliated Hospital of Jinzhou Medical University, Jinzhou 121000,China;Chinese PLA General Hospital, Beijing 100853,China;Beijing Institute of Biotechnology, Beijing 100850,China)

机构地区：[1]锦州医科大学附属第一医院,辽宁锦州121000 [2]军事医学科学院生物工程研究所,北京100850 [3]解放军总医院,北京100853

出　　处：《生物技术通讯》2017年第5期610-613,714,共5页Letters in Biotechnology

基　　金：国家自然科学基金(81672602;81472589;81502264);北京市科技新星计划(Z141102001814055);军事医学科学院创新基金转化医学项目(ZHYX003)

摘　　要：目的:构建携带Myc标签的人细胞外信号调节激酶2(ERK2)的真核表达载体,表达Myc-ERK2融合蛋白,检测其对人视网膜母细胞瘤细胞生长的影响。方法:采用PCR技术从乳腺文库中扩增人ERK2基因序列,将其插入pXJ-40载体;将重组质粒与空载体转染人视网膜母细胞瘤WERI-Rb-1细胞系后,Western印迹检测表达情况,并进行生长曲线实验。结果:双酶切和测序鉴定表明Myc-ERK2真核表达质粒构建成功;转染WERI-Rb-1细胞后融合蛋白获得表达,生长曲线实验结果表明ERK2可促进肿瘤细胞的生长。结论:携带Myc标签的人ERK2基因的真核表达载体能在人视网膜母细胞瘤WERI-Rb-1细胞中表达,且能促进该细胞的生长。本实验为进一步研究ERK2在肿瘤尤其是眼恶性肿瘤中的功能奠定了基础。Objective:To construct the eukaryotic expression vector of human extracellular signal regulated kinase2(ERK2)labeled with Myc and detect its expression and activity in human retinoblastoma cells.Methods:Human ERK2coding region was amplified from human mammary cDNA library by PCR and cloned into pXJ-40vector.The recombinant plasmid Myc-ERK2was transfected into human WERI-Rb-1cells and the expressionwas detected by Western blot.The proliferation of WERI-Rb-1cells was examined by growth curve experiment.Results:Myc-ERK2eukaryotic expression vector was constructed and verified by double enzyme digestion and nucleic acid sequencing.Human ERK2protein was expressed in human WERI-Rb-1cells as was shown by Western blot results and ERK2protein promoted the proliferation of WERI-Rb-1cells.Conclusion:The Myc-ERK2was identified to promote WERI-Rb-1cell proliferation,which is a fundamental study for ERK2's role in retinob-laastoma.

关 键 词：细胞外信号调节激酶2(ERK2) 真核表达 人视网膜母细胞瘤细胞 

分 类 号：Q78[生物学—分子生物学]