EphA3促进胃癌细胞BGC823的增殖  

EphA3 Promotes the Proliferation of Gastric Cancer Cell BGC823

作　　者：吕晓业 王健[2] 李昕宇[3] 黄芳[2] 王芃[2] 周建光[2] 李山虎[2] 卫勃[1] 

机构地区：[1]解放军总医院普外科,北京100853 [2]军事医学科学院生物工程研究所,北京100850 [3]湖北医药学院生物医学工程学院,湖北十堰442000

出　　处：《生物技术通讯》2017年第3期286-289,共4页Letters in Biotechnology

基　　金：国家自然科学基金(81372140;81372770;81470138)

摘　　要：目的:初步探索EphA3与胃癌细胞BGC823增殖的关系。方法:用T_4DNA连接酶将EphA3的全长cDNA片段连接到酶切后的慢病毒载体pCDH-EF1-MCS-T2A-Puro中制备慢病毒,感染胃癌BGC823细胞建立过表达EphA3细胞系,Western印迹检测目的蛋白的表达,CCK8细胞生长实验和裸鼠成瘤实验检测EphA3对BGC823细胞增殖的影响。结果:双酶切鉴定与基因测序表明过表达EphA3慢病毒载体构建成功;Western印迹表明建立了过表达EphA3的BGC823细胞系,高表达EphA3对BGC823细胞的增殖和成瘤后生长有明显的促进作用。结论:EphA3促进BGC823胃癌细胞增殖,为胃癌的靶向性治疗及个性化治疗提供了线索。Objective： To explore the function of EphA3 on cell proliferation in BGC823 cells. Methods： T4 DNA ligase was used to ligate exogenous cDNA fragment of EphA3 into the pCDH-EF1-MCS-T2A-Puro vector. The recombinant plasmid with lentivirus packaging plasmids was transfeeted into 293T cells lines and lentiviral particles were collected at different time. BGC823 cells were infected with lentiviral particles and expression of EphA3 was verified by Western blotting. Moreover, CCK8 test and xenograft assay in nude mice were used to examine the proliferation of the infected BGC823 cells with EphA3 overexpression. Results： Double enzyme digestion and gene sequencing proved that the construction of plasmid was successful. The lentiviral plasmid can upregulate the EphA3 protein level in BGC823 cells. EphA3 overexpression promotes gastric cancer BGC823 ceils proliferation in vitro and in vivo. Conclusion： In BGC823 ceils, EphA3 overexpression can promote tumor cell proliferation, and perhaps play an important role in gastric cancer development.

关 键 词：EphA3 胃癌 BGC823细胞 

分 类 号：Q78[生物学—分子生物学] Q25