沉默SEMA3A基因对胶质瘤细胞U251增殖和转移的影响

探讨了SEMA3A基因在恶性胶质瘤细胞U251迁移和侵袭能力中的作用, 及其用于胶质瘤临床治疗的可行性. 用特异识别SEMA3A基因的短发卡RNA(short hairpin RNA,shRNA)片段与真核表达载体pFH-GFP-L连接, 再与辅助质粒联用来包装慢病毒. 通过荧光显微镜观察感染胶质瘤细胞U251的感染效率. 通过实时定量PCR技术和Western-blot技术验证了U251细胞中SEMA3A基因的沉默效果. 通过MTT法、PI染色法和Transwell小室分别检测了U251细胞增殖、细胞周期和运动能力的变化. 结果表明, SEMA3A-shRNA慢病毒感染U251能有效下调SEMA3A基因的表达水平(P <0.01), 使U251细胞的增殖能力、迁移和侵袭能力明显降低(P <0.05), 细胞大量阻滞在G2/M期, 并促进了细胞凋亡(P <0.05).

Effect of RNA interference targeting SEMA3A on proliferation and metastasis of glioma cell line U251

This paper explores the effect of knockdown SEMA3A gene on the migration and invasion of glioma cell line U251. Preliminary discussion of the potential in treatment of glioma was carried out. The highest knockdown efficiency plasmid packaged by a lentivirus packaging vector system was constructed, which contained a pFH-GFP-L vector and a short hairpin RNA (shRNA) sequence targeting the human SEMA3A gene. U251 cells were infected with a modified lentivirus vector. The infection efficiency of shRNA targeting SEMA3A gene was observed by fluorescence microscope. Real-time PCR reaction and
Western-blot were used to investigate the expression of mRNA and protein of SEMA3A. The results show that the lentivirus containing SEMA3A target shRNA are efficient in silencing SEMA3A expression (P < 0.01). Knockdown SEMA3A gene expression could significantly inhibit the proliferation and metastasis of U251 cells (P < 0.05). In addition,a large number of U251 cells are arrested in G2/M phase, promoting apoptosis (P < 0.05).