siRNA干扰PDGFRβ抑制新生儿血管瘤干细胞的增殖和分化

siRNA Inhibits Proliferation and Differentiation of Infant Hemangioma Stem Cells by Interfering with PDGFRβ

为了研究血小板衍生生长因子受体β(PDGFRβ)在血管瘤干细胞(Hem-SCs)中的表达,并探讨PDGFRβsiRNA对Hem-SCs增殖、凋亡和分化的影响。本研究通过组织消化法和免疫磁珠分选法从新生儿血管瘤组织中分离Hem-SCs,并通过流式细胞术鉴定细胞免疫表型;并使用Western blotting法检测PDGFRβ蛋白在Hem-SCs中的表达水平。通过lipofectamine 2000将PDGFRβsiRNA转染到Hem-SCs细胞,使用CCK8和流式细胞术分别检测PDGFRβsiRNA对Hem-SCs增殖和凋亡的影响;经过Western blotting检测PDGFRβsiRNA对Hem-SCs Sox2、Oct4和Notch1蛋白表达的影响。流式细胞术检测结果显示,CD133和CD90在分离的Hem-SCs中阳性率分别为99.8%和96.7%;Western blotting结果显示PDGFRβ蛋白在Hem-SCs细胞中的表达显着高于血管瘤内皮细胞;CCK8结果显示,与Control组比较,PDGFRβsiRNA组细胞增殖率显著降低(p<0.01);流式检测结果显示,与Control组比较,PDGFRβsiRNA组Hem-SCs凋亡率为显著升高(p<0.01);Western blotting结果显示,与Control组比较,PDGFRβsiRNA组Sox2、Oct4和Notch1蛋白表达显著降低(p<0.01)。本研究结果表明,siRNA干扰PDGFRβ能够抑制Hem-SCs增殖和分化,促进Hem-SCs凋亡。

To study the expression of platelet derived growth factor receptor β(PDGFRβ) in infantile hemangioma stem cells(Hem-SCs) and to investigate the effect of PDGFRβ si RNA on proliferation, apoptosis and differentiation of Hem-SCs. Hem-SCs were isolated from hemangiomas by tissue digestion and immunomagnetic bead sorting,and the cellular immune-phenotype was identified by flow cytometry. The expression of PDGFRβ protein in Hem-SCs were detected by Western blotting. PDGFRβ siRNA was transfected into Hem-SCs cells by Lipofectamine 2000. The effects of PDGFRβ siRNA on the proliferation and apoptosis of Hem-SCs were detected by CCK8 and flow cytometry. Western blotting was used to detect the expression of Sox2, Oct4 and Notch1 proteins in Hem-SCs transfected with PDGFRβ siRNA. The results of flow cytometry showed that the positive rates of CD133 and CD90 in isolated Hem-SCs were 99.8% and 96.7% respectively. Western blotting results showed that the expression of PDGFRβ protein in Hem-SCs was significantly higher than that in hemangioma epidermal cells.The results of CCK8 showed that the proliferation rate of Hem-SCs in PDGFRβ siRNA transfection group was significantly lower than that in the control group(p<0.01). The results of flow cytometry showed that the apoptosis rate of Hem-SCs cells in the PDGFRβ siRNA group was significantly higher than that of the control group(p<0.01).Western blotting results showed that the expression of Sox2, Oct4 and Notch1 in the PDGFR mrna group was significantly decreased compared with the control group(p<0.01). The results of this study showed that interference with PDGFRβ by siRNA can inhibit the proliferation and differentiation of Hem-SCs and promote the apoptosis of Hem-SCs.