USP14全长及截短表达质粒构建及其在子宫内膜癌中功能的初探  

Construction of USP14 full-length and truncated expression plasmid and its function in endometrial cancer

作　　者：苏英杰 曾凯[2] 赵越[2] 张颐[1] SU Ying-jie;ZENG Kai;ZHAO Yue;ZHANG Yi(Department of Cynerology,First Affiliated Hospital of China Medical University,Shenyang 110001;School of Life Sciences,China Medical Universitv,Key Laboratory of Cell Biology,Ministry of Health,Key Laboratory of Medical Cell Biology,Ministry of Education,Laboratory of Chromatin Biology,Shenyang 110122,China)

机构地区：[1]中国医科大学附属第一医院妇科,辽宁沈阳110001 [2]中国医科大学生命科学学院卫生部细胞生物学重点实验室,教育部医学细胞生物学重点实验室,染色质生物学研究室,辽宁沈阳110122

出　　处：《解剖科学进展》2021年第4期385-388,392,共5页Progress of Anatomical Sciences

基　　金：辽宁省中央引导地方科技发展专项(2019JH6/10400006);中国医科大学2019年度临床医学培育学科支持计划(妇产科学-机器人);中国医科大学国际水平项目(2019年);国家重点研发计划项目(2018YFC1311600)。

摘　　要：目的构建USP14全长及截短表达质粒并且初步探讨USP14在子宫内膜癌中的作用。方法设计USP14全长及截短的特异性引物,通过PCR合成相应cDNA片段,并用限制性内切酶EcoR I和Xho I进行双酶切,连接转化后将构建好的质粒进行测序,成功构建后将其转染至COS7中,观察全长及截短在COS7中分布。通过划痕实验以及MTS实验探究其对子宫内膜癌细胞生物学功能的影响。结果成功构建USP14全长及截短表达质粒,在COS7细胞中,全长及C端截短定位于细胞浆中,N端截短主要定位于细胞核;USP14能够促进子宫内膜癌细胞的迁移并提高子宫内膜癌细胞的生存活力。结论在COS7细胞中,USP14全长及C端截短分布于细胞浆,N截短大部分分布于细胞核,USP14能够促进子宫内膜癌细胞的生长和迁移。Objective To construct the full-length and truncated expression plasmids of USP14 and preliminarily explore the role of USP14 in endometrial cancer. Methods Design specific primers for the full-length and truncated USP14, synthesize corresponding cDNA fragments by PCR, and use restriction enzyme EcoR Ⅰ double-enzyme digestion with Xho Ⅰ. After ligation and transformation, the constructed plasmid will be sequenced. After successful construction, it will be transfected into COS7. Observe the distribution of full length and truncation in COS7. Through the scratch experiment and MTS experiment to explore its effect on the biological function of endometrial cancer cells, the USP14 full-length and truncated expression plasmids were successfully constructed. Results In COS7 cells, the full-length and C-terminal truncations are located in the cytoplasm. The N-terminal truncation is mainly located in the nucleus;USP14 can promote the migration of endometrial cancer cells and improve the viability of endometrial cancer cells. Conclusion In COS7 cells, the full length and C-terminal truncation of USP14 are distributed in the cytoplasm. The shortest part is distributed in the nucleus. USP14 can promote the proliferation and migration of endometrial cancer.

关 键 词：USP14 截短 子宫内膜癌 细胞活力 迁移 

分 类 号：R737.33[医药卫生—肿瘤]