Syndecan-1基因沉默抑制胶质瘤A172细胞增殖和侵袭  

Syndecan-1 knockdown inhibits the proliferation and invasion of A172 glioblastoma multiforme cells

作　　者：石爽[1] 钟东[1] 王兵[1] 王文涛[1] 张福安[1] 黄浩洋[1] 

机构地区：[1]重庆医科大学附属第一医院神经外科,重庆医科大学附属第一医院实验研究中心,重庆400016

出　　处：《中国神经精神疾病杂志》2016年第2期74-79,共6页Chinese Journal of Nervous and Mental Diseases

基　　金：重庆市科委自然科学基金科研项目(编号:cstc2011jj A10091);财政部;卫生部国家临床重点专科建设项目[编号:财社(2001)170号]

摘　　要：目的探讨多配体蛋白多糖-1(Syndecan-1,SDC1)在不同胶质瘤细胞株中的表达水平及其沉默对A172细胞的增殖和侵袭的影响。方法通过实时荧光定量PCR(q RT-PCR)和Western Blotting分析SDC1在不同胶质瘤细胞株中的表达水平;将携带SDC1 sh RNA的慢病毒载体感染A172细胞,稳定沉默的细胞株为干扰组,未转染为阴性对照组,转染scramble序列为空白对照组;采用MTT法、台盼蓝拒然法和流式细胞术检测细胞增殖能力,Transwell小室实验检测细胞的迁移和侵袭力;q RT-PCR和Western Blotting检测相关蛋白变化。结果SDC1在不同的胶质瘤细胞株中表达强度不同,差异具有统计学意义(P<0.05)。成功构建稳定沉默SDC1的A172细胞株;与阴性对照组和空白对照组相比,干扰组细胞增殖受到抑制(P<0.05),迁移力(58.40±5.24 vs.255.8±16.09、226.5±22.84,F=126.4,P<0.05)和侵袭力(61.67±16.26 vs.233.70±17.24、244.30±28.15,F=69.87,P<0.05)明显抑制;SDC1、PCNA和MMP-9 m RNA和蛋白表达水平明显降低(P<0.05)。结论沉默SDC1基因的表达可抑制胶质瘤A172细胞的增殖、迁移和侵袭,提示SDC1可能成为胶质瘤生物治疗的新靶点。Objective To investigate the expression of syndecan-1 （SDC 1） in glioma cells and the effects of synde- can-1 knockdown on the proliferation and invasion of A172 cells. Methods The expression of syndecan-1 in glioma cells was analyzed using quantitative Real-time PCR and Western blotting. A172 cells were transfected with lentiviral vector carrying SDC1 shRNA to establish a stable SDCl-silencing cell line. The cell proliferation was analyzed by MTY assay. Trypan blue exclusion assay and flow cytometry, and Transwell assays were performed to measure the migration and invasion abilities, respectively. The mRNA and protein and expression levels of SDC1, Proliferation Cell Nuclear An- tigen （PCNA） and Matrix Metalloproteinase 9 （MMP-9） were detected by using qRT-PCR and Western blotting. Results The expression levels of SDC1 were significantly different in different glioma cell lines. The stable SDCl-silencing cell line was successfully established, in which the mRNA and protein expression levels of SDC1 were significantly decreased （P 〈 0.05）. SDC1 knockdown significantly reduced the cell proliferation, migration（58.40±5.24 vs. 255.8±16.09,226.5± 22.84, F=126.4, P 〈 0.05 ） and invasion （61.67 ± 16.26 vs. 233.70± 17.24,244.30±28.15, F=69.87, P 〈 0.05） compared with either control group or blank group. SDC1 knockdown also significantly decreased the mRNA and protein expression levels of PCNA and MMP-9 （P 〈 0.05）. Conclusion： SDC1 knockdown suppresses the capacities of proliferation, invasion and migration of glioma A 172 cell, implying that SDC 1 may serve as a novel target in the biotherapy of glioma.

关 键 词：胶质瘤 SYNDECAN-1 增殖和侵袭 

分 类 号：R739.41[医药卫生—肿瘤]