过表达CSRP1基因可促进人肺腺癌H1299细胞的增殖、迁移和侵袭  

Overexpression of CSRP 1 gene promotes proliferation, migration and invasion of human lung adenocarcinoma H1299 cells

作　　者：张笑 余涛 林河春[1] 耿沁[1] 潘洪玉 姚明[1] ZHANG Xiao;YU Tao;LIN Hechun;GENG Qin;PAN Hongyu;YAO Ming(State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, Chin)

机构地区：[1]上海交通大学医学院附属仁济医院上海市肿瘤研究所癌基因与相关基因国家重点实验室,上海200032

出　　处：《肿瘤》2018年第4期291-299,共9页Tumor

基　　金：国家自然科学基金资助项目(编号:81702846);上海市卫生和计划生育委员会科研计划资助项目(编号:20174Y0183)~~

摘　　要：目的:探讨半胱氨酸和甘氨酸丰富蛋白1(cysteine and glycine rich protein 1,CSRP 1)基因过表达对人肺腺癌H1299细胞增殖、周期、迁移和侵袭的影响,及其可能的作用机制。方法:构建携带有CSRP 1基因的重组慢病毒质粒pCDH-CSRP1;将重组慢病毒Ad-CDH-CSRP1和pCDH-GFP(空载体对照组)分别感染人肺腺癌H1299细胞。实时荧光定量PCR及蛋白质印迹法检测CSRP1 mRNA及蛋白在H1299细胞中的表达水平。CCK-8法和克隆形成实验检测CSRP 1基因过表达对H1299细胞增殖能力的影响;FCM法检测CSRP 1基因过表达对H1299细胞周期的影响。Transwell小室迁移实验和划痕愈合实验检测CSRP 1基因过表达对H1299细胞纵向和横向迁移能力的影响,Transwell小室侵袭实验检测对细胞侵袭能力的影响。蛋白质印迹法检测CSRP 1基因过表达对H1299细胞中黏着斑激酶(focal adhesion kinase,FAK)和磷酸化FAK(phospho-FAK,p-FAK)蛋白表达水平的影响。结果:重组慢病毒质粒pCDH-CSRP1构建成功。转入重组慢病毒质粒pCDH-CSRP1的H1299细胞中CSRP1蛋白的表达水平较pCDH-GFP组明显提高(P<0.01)。CSRP 1基因过表达可促进H1299细胞增殖(P<0.05),G1期细胞所占百分比明显下降(P<0.01),S期细胞所占百分比明显上升(P<0.01);CSRP 1基因过表达可明显增强H1299细胞的迁移和侵袭能力(P值均<0.01)。CSRP 1基因过表达的H1299细胞中p-FAK蛋白的表达水平明显升高(P<0.01),而总FAK蛋白的表达水平无明显变化(P>0.05)。结论:CSRP 1基因过表达可增强人肺腺癌H1299细胞的增殖、迁移和侵袭能力,其机制可能与FAK信号转导通路的激活有关。Objective： To investigate the effects of cysteine and glycine rich protein 1 （CSRP1） gene overexpression on proliferation, cell cycle, migration and invasion of human lung adenocarcinoma H1299 cells, and to explore the posssible mechanism.Methods： The recombinant lentivirus plasmids pCDH-CSRP1 carrying CSRP1 gene and pCDH- GFP （as the control） were constructed and infected into human lung adenocarcinoma H1299 cells, respectively. The expression levels of CSRP1 mRNA and protein in H1299 cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The proliferation of H1299 cells with CSRP1 gene overexpression was tested by CCK-8 assay and colony formation assay. The cell cycle distribution of H1299 cells was detected by FCM. The longitudinal and lateral migration abilities of H1299 cells were detected by Transwell chamber assay and scratch wound healing assay. The invasion ability of H1299 cells was detected by Transwell chamber assay. The expression levels of focal adhesion kinase （FAK） and phospho-FAK （p-FAK） proteins in H1299 cells were detected by Western blotting.Results： The recombinant lentivirus plasmid pCDH-CSRP1 was successfully constructed, and the expression of CSRP1 in H1299 cells transfected with pCDH-CSRP1 was significantly higher than that in H1299 cells transfected with pCDH-GFP （P 〈 0.01）. The overexpression of CSRP1 gene promoted the proliferation of H1299 cells （P 〈 0.05）, and contributed to the increased proportion of G1 phase （P 〈 0.01） and the decreased proportion of S phase （P 〈 0.01） in H1299 cells. Furthermore, the overexpression of CSRP1 gene facilitated the migration and invasion abilities of H1299 cells （both P 〈 0.01）. Compared with the control group, the expression level of p-FAK increased significantly （P 〈 0.01）, while the expression level of FAK remained unchanged （P 〉 0.05） in H1299 cells with CSRP1 gene overexpression.Conclusion： CSRP1 gene overexpression can promote the prolif

关 键 词：肺肿瘤 细胞增殖 细胞运动 细胞周期 CSRP1 基因 

分 类 号：R737.9[医药卫生—肿瘤]