摘要
目的探讨趋化因子CXCL12受体CXCR4 sh RNA对胶质瘤细胞U87增殖及血管内皮生长因子(vascular endothelial growth factor,VEGF)和白细胞介素-8(interleukin-8,IL-8)表达的影响,为胶质瘤的基因治疗提供实验依据。方法用脂质体Lipofectimine TM 2000分别将CXCR4 sh RNA载体p GPU6/GFP/Rac1-421和非特异质粒p GPU6/GFP/NC转染U87细胞,并设未转染组,转染后48 h,采用RT-PCR法检测各组细胞中CXCR4基因m RNA的转录水平;转染后24、48、72 h,采用MTT法检测细胞的增殖活力,ELISA法检测细胞培养上清中VEGF和IL-8的含量。结果转染p GPU6/GFP/Rac1-421可使U87细胞中CXCR4基因m RNA的转录水平、细胞体外增殖活力及细胞培养上清中VEGF和IL-8的含量均明显降低,与p GPU6/GFP/NC组和未转染组比较,差异有统计学意义(P<0.05)。结论 CXCR4 sh RNA可下调U87细胞中CXCR4基因m RNA的转录,抑制细胞增殖,降低VEGF和IL-8的产生,提示CXCR4在胶质瘤发展过程中具有重要作用。
Objective To investigate the effect of CXL12 receptor CXCR4 sh RNA on the proliferation of glioma U87 cells and expressions of vascular endothelial growth factor(VEGF)and interleukin-8(IL-8) and provide an experimental basis for gene therapy of glioma. Methods U87 cells were transfected with CXCR4 sh RNA vector p GPU6 / GFP / Rac1-421 and non-specific plasmid p GPU6 / GFP / NC in mediation of Lipofectimine^TM 2000 respectively,using those untransfected as control. The transcription levels of CXCR4 m RNA in cells of various groups were determined by RT-PCR 48 h after transfection. The proliferative activity of cells was determined by MTT method 24,48 and 72 h after transfection,while the VEGF and IL-8 contents in cell culture supernatant by ELISA. Results The CXCR4 m RNA transcription level,in vitro proliferative activity,as well as VEGF and IL-8 contents in cell culture supernatant of U87 cells transfected with p GPU6 / GFP / Rac1-421 were significantly lower than those in the cells transfected with p GPU6 / GFP / NC and untransfected(P〈0. 05). Conclusion CXCR4 sh RNA down-regulated the transcription of CXCR4 m RNA in U87 cells,inhibited the cell proliferation,and decreased the production of VEGF and IL-8,which indicated that CXCR4 played an important role in the development process of glioma.
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第12期1539-1542,共4页
Chinese Journal of Biologicals