miRNA-126表达阻断与成釉细胞瘤细胞血管生成相关因子表达水平的关系

Relationship between miRNA-126expression blocking and the expression level of angiogenesis related factors in ameloblastoma cells

目的研究miRNA-126表达阻断后,成釉细胞瘤内血管的变化情况。方法将实验对象分为Control组(未转染组),NC组(转染无意义单链反义核苷酸药物的细胞组),AMO组(转染单链反义核苷酸药物的细胞组)。通过制备miRNA-126的单链反义核苷酸药物,加入到体外培养的成釉细胞瘤细胞中,荧光定量PCR检测体外培养的成釉细胞瘤细胞中miRNA-126表达是否被阻断。利用荧光定量PCR和Western blot分别在RNA和蛋白水平检测血管内皮生长因子(vascular endothelial growth factor,VEGFA)、细胞间黏附因子-1(intercellular adhesion molecule-1,ICAM-1)(CD54)、血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)(CDl06)等相关细胞因子的表达情况。结果与Control组相比,AMO组miRNA-126表达水平显著降低,差异极显著,而VEGFA、ICAM-1和VCAM-1的RNA表达水平及蛋白表达水平显著升高。miRNA-126的单链反义核苷酸药物可以有效的阻断miRNA-126的表达,miRNA。126表达阻断后,成釉细胞瘤细胞中VEGFA和成纤维细胞生长因子7(fibroblast growth factor 7,FGF7)表达显著升高。结论miRNA-126的单链反义核苷酸可能会通过阻断miRNA-126的表达,进而增加VEGFA、ICAM-1和VCAM-1的表达,从而达到调节成釉细胞瘤生长的目的。

Objective To investigate the changes of angiogenesis in ameloblastoma after blocking the expression of miRNA-126.Methods This experiment is divided into 3groups.Control(untransfeeted cells),NC group (cells transfected with scramble single-stranded antisense nucleotide),AMO group (cells transfected with single-stranded antisense nueleotides).The single-stranded antisense oligonucleotides of miRNA-126were pre- pared and added to cultured ameloblastoma cells in vitro.The expression of miRNA-126in ameloblastoma cells was detected by fluorescence quantitative PCR.Fluorescence quantitative PCR and Western Blot were used to detect the expression of vascular endothelial growth factor(VEGFA),intercellular adhesion molecule-1(ICAM- 1)and vascular cell adhesion molecule-1(VCAM-1)in RNA and protein levels,respectlvely.Results Compared with the control cells,the expression of miRNA-126 in AMO group was significantly lower.The mRNA and protein levels of VEGFA,ICAM-1 and VCAM-1 in AMO group were significantly increased.Single-stranded antisense ollgonucleotide of miRNA-126can effectively block miRNA-126.The expression of VEGFA and FGF7in ameloblastoma cells was significantly increased after the expression of -126miRNA was blocked.Conclusions Single-stranded antisense oligonucleotides of miRNA-126increased the expression of VEGFA,ICAM-1and VCAM-1via blocking the expression of miRNA-126.