RSPO2过表达人肝星状细胞株LX-2中差异表达蛋白的筛选及生物学功能分析  

Screening differential proteins of human hepatic stellate cell line LX-2 with over-expressed RSPO2 and biological function analysis for the differentially expressed proteins

作　　者：殷新光 薛文英 徐英 吴一鸣[2] 袁芳 邹丹丹 YIN Xinguang;XUE Wenying;XU Ying;WU Yiming;YUAN Fang;ZOU Dandan(The Affiliated Women and Children′s Hospital of Jiaxing College, Jiaxing 314000, China)

机构地区：[1]嘉兴学院附属妇女儿童医院,浙江嘉兴314000 [2]嘉兴学院附属第一医院 [3]杭州佰辰医学检验所有限公司

出　　处：《山东医药》2019年第15期1-5,共5页Shandong Medical Journal

基　　金：浙江省自然科学基金(LY16H030016)

摘　　要：目的 筛选特异性顶部盘状底板反应蛋白2(RSPO2)在人肝星状细胞株LX-2中过表达后的差异表达蛋白,并进行差异表达蛋白的生物学功能分析。方法 通过分子生物学手段构建RSPO2过表达载体并转染进人肝星状细胞,利用RT-PCR验证RSPO2的过表达状态,细胞划痕实验观察过表达RSPO2肝星状细胞的生长及迁移能力。通过液相色谱-串联质谱(LC-MS/MS)技术检测RSPO2过表达人肝星状细胞和正常人肝星状细胞的差异表达蛋白,并对差异表达蛋白进行生物信息学分析。结果 通过RT-PCR验证,在人肝星状细胞株LX-2中成功过表达了RSPO2蛋白;通过LC-MS/MS技术对比RSPO2过表达肝星状细胞和正常肝星状细胞,获得候选差异表达蛋白共计233个,其中115个蛋白在RSPO2过表达细胞中表达下调,118个表达上调,这些差异表达蛋白参与了很多与细胞转录、复制、增殖相关的信号通路。结论 RSPO2过表达LX-2细胞差异表达蛋白共计233个,其中115个蛋白在RSPO2过表达细胞中表达下调,118个表达上调,这些差异表达蛋白大多参与到RNA的转录与翻译过程。Objective To screen out the differential proteins of human hepatic stellate cells LX-2 with over-expressed roof plate-specific spondin 2 (RSPO2), and to analyze the biological function of the differentially expressed proteins. Methods The RSPO2 overexpression vector was constructed by molecular biology and transfected into human hepatic stellate cells. The overexpression status of RSPO2 was verified by RT-PCR, and the cell Scratch assay was used to observe the growth and migration abilities. The differential proteins between human hepatic stellate cells with over-expressed RSPO2 and normal human hepatic stellate cells were detected by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) technique, and the differential proteins were analyzed by bioinformatics analysis. Results The RSPO2 gene was successfully overexpressed in human hepatic stellate cell line LX-2, which was verified by RT-PCR. The differential proteins in the human hepatic stellate cells with overexpression of RSPO2 and normal human hepatic stellate cells were compared by LC-MS/MS. A total of 233 candidate differential proteins were obtained, of which, 115 proteins in the RSPO2 overexpressed cells were down-regulated and 118 proteins were up-regulated. These differential proteins were involved in many signaling pathways which were related to cell transcription, replication, and proliferation. Conclusion A total of 233 candidate differential proteins in LX-2 cells with over-expressed RSPO2 are obtained, of which, 115 proteins are down-regulated and 118 proteins are up-regulated, and most of these differential proteins are involved in the transcription and translation of RNA.

关 键 词：特异性顶部盘状底板反应蛋白2 肝星状细胞 差异表达蛋白 

分 类 号：R735.7[医药卫生—肿瘤]