摘要
目的构建带myc标签的人Yes相关转录调节蛋白1(YAP1)基因的真核表达载体,探讨YAP1对甲状腺癌BCPAP细胞增殖和迁移的影响。方法采用聚合酶链反应(PCR)扩增YAP1的编码区序列,将其插入到pXJ40-myc载体中,构建pXJ40-myc-YAP1重组质粒并鉴定;质粒转染人甲状腺癌BCPAP细胞,Western印迹鉴定融合蛋白表达,细胞增殖实验(CCK-8法)、克隆形成实验、划痕实验检测细胞的生长、增殖和迁移能力的变化。结果 pXJ40-mycYAP1质粒构建成功;转染BCPAP细胞后重组融合蛋白表达成功;细胞增殖实验、克隆形成及划痕实验证明YAP1可促进甲状腺癌细胞BCPAP的增殖和迁移。结论带myc标签的人YAP1基因能在人甲状腺癌BCPAP细胞中表达,且可促进该细胞的增殖和迁移,为进一步研究YAP1在甲状腺肿瘤中的功能奠定了基础。
Objective To construct the gene eukaryotic expression vector of human Yes related protein 1(YAP1)with myc tag and investigate the effect of YAP1 expression in BCPAP cells on proliferation and migration. Methods The coding region of YAP1 was amplified by PCR and inserted into the pXJ40-myc vector to construct the recombinant plasmid pXJ40-myc-YAP1. The recombinant plasmid was transfected into human thyroid cancer BCPAP cells,and the expression of fusion protein was identified by Western blotting. The growth,proliferation and migration of cells were detected by cell proliferation assay(CCK-8 method),clone formation assay and wound-healing assay. Results The plasmid pXJ40-mycYAP1 was constructed. After transfection of BCPAP cells,the recombinant fusion protein was expressed. YAP1 could promote BCPAP proliferation and migration of thyroid cancer cells as evidenced by cell proliferation,clone formation and wound-healing assay. Conclusion Human YAP1 gene carrying myc tag can be expressed in human thyroid cancer BCPAP cells and promote proliferation and migration of these cells. which can contribute to the study of the function of YAP1 in human thyroid cancer cells,especially thyroid tumors.
作者
康伟
王晓侠
刘丹
张鹏飞
KANG Wei;WANG Xiao-xia;LIU Dan;ZHANG Peng-fei(Department of Otolaryngology Head and Neck Surgery,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;Department of Otolaryngology Head and Neck Surgery,Air Force 986 Hospital,Xi'an 710054,China;Department of Cardiovascular Surgery,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China)
出处
《军事医学》
CAS
2021年第9期675-678,695,共5页
Military Medical Sciences
基金
国家自然科学基金青年科学基金项目(81602562)
陕西省国际科技合作与交流计划(2017KW-063)
西安交通大学校基金(1191329722)
西安交通大学第一附属医院院基金(2015YK17)。
关键词
Yes相关蛋白1
甲状腺癌
BCPAP细胞
真核表达
细胞生长
细胞迁移
Yes1 associated transcriptional regulator
thyroid cancer
BCPAP cell
eukaryotic expression
cell growth
cell migration