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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 effects of chemokine CXCL12 receptor CXCR4 sh RNA on the proliferation of glioma U87 cells and the expression of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) Provide experimental evidence 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 respectively using lipofectamine 2000. After 48 h, The transcript levels of CXCR4 mRNA in each group were detected by RT-PCR. At 24, 48 and 72 h after transfection, the proliferation activity of the cells was detected by MTT assay. The levels of VEGF and IL-8 Content. Results Transfection of p GPU6 / GFP / Rac1-421 could significantly decrease the transcription level of mCRNA, the activity of cell proliferation and the content of VEGF and IL-8 in the cell culture supernatant of U87 cells, / NC group and untransfected group, the difference was statistically significant (P <0.05). Conclusion CXCR4 sh RNA can down-regulate the transcription of CXCR4 m RNA in U87 cells, inhibit cell proliferation and decrease the production of VEGF and IL-8, suggesting that CXCR4 plays an important role in the development of glioma.