石爽钟东王兵王文涛张福安黄浩洋

Syndecan-1基因沉默抑制胶质瘤A172细胞增殖和侵袭☆

石爽*钟东*王兵*王文涛*张福安*黄浩洋*

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（qRT-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 shRNA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；qRT-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 mRNA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。

胶质瘤 Syndecan-1增殖和侵袭

【Abstract】Objective To investigate the expression of syndecan-1（SDC1）in glioma cells and the effects of syndecan-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 SDC1-silencing cell line.The cell proliferation was analyzed by MTT 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 Antigen（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 SDC1-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 expressionlevels of PCNA and MMP-9（P＜0.05）.Conclusion：SDC1 knockdown suppresses the capacities of proliferation，invasion and migration of glioma A172 cell，implying that SDC1 may serve as a novel target in the biotherapy of glioma.

【Key words】Glioma Syndecan-1 Proliferation and Invasion

脑胶质瘤是中枢神经系统最常见的原发性颅内肿瘤，患者的中位生存期低于15个月，目前临床综合治疗虽然取得一定进展，但其疗效并不理想［1］。多配体蛋白多糖-1（Syndecan-1，SDC1）是一类Ⅰ型跨膜蛋白多糖，可与广泛的胞外配体及胞膜受体相结合［2］，介导胞内外信号的传导，与肿瘤的发生和进展密切相关［3］。然而SDC1在胶质瘤上是否表达尚不确定［4，5］。本课题在此基础上检测SDC1在不同胶质瘤细胞株中的表达水平，利用RNAi技术研究SDC1对胶质瘤增殖和侵袭的影响，寻找胶质瘤生物治疗的新靶点。……