甲基硒酸对婴幼儿血管瘤内皮细胞增殖和凋亡的影响及其分子机制的研究

Effect of Methylselenic Acid on Proliferation and Apoptosis of Endothelial Cells in Infantile Hemangioma and Its Molecular Mechanism

目的探讨甲基硒酸(MSA)对婴幼儿血管瘤内皮细胞增殖、凋亡的影响及其分子机制。方法采用不同浓度MSA作用于血管瘤内皮细胞24 h,MTT法检测细胞增殖抑制率,流式细胞仪检测细胞凋亡,Western-blot检测增殖、凋亡相关蛋白的表达,实时荧光定量PCR(RT-q PCR)检测miR-499a-5p的表达。转染miR-499a-5p模拟物及其抑制物至血管瘤内皮细胞,采用上述方法检测过表达miR-499a-5p及抑制miR-499a-5p表达对血管瘤内皮细胞增殖和凋亡的影响。结果 MSA作用于血管瘤内皮细胞后,细胞增殖抑制率、凋亡率、miR-499a-5p表达显著升高,且具有显著的量效关系(P<0.05)。过表达miR-499a-5p后血管瘤内皮细胞增殖抑制率、凋亡率显著升高(P<0.05)。抑制miR-499a-5p表达可部分逆转MSA对血管瘤内皮细胞增殖和凋亡的影响(P<0.05)。结论 MSA可抑制婴幼儿血管瘤内皮细胞增殖,促进细胞凋亡,其机制可能与上调miR-499a-5p表达有关。

Objective To explore the effect of methylselenic acid(MSA)on proliferation and apoptosis of endothelial cells in infantile hemangiomas and its molecular mechanism.Methods Hemangioma endothelial cells were treated with different concentrations of MSA for 24 hours,the methyl thiazolyl tetrazolium(MTT)method was used to detect cell proliferation inhibition rate,flow cytometry was used to detect apoptosis,Western-blot was used to detect the expression of proliferation and apoptosis related proteins,and real-time fluorescence quantitative PCR(RT-qPCR)was applied to detect expression level of miR-499a-5p.miR-499a-5p mimics and anti-miR-499a-5p were transfected into hemangioma endothelial cells,and the effect of overexpression and inhibition of miR-499a-5p on proliferation and apoptosis of hemangioma endothelial cells were detected by the above methods.Results After being treated by MSA,the cell proliferation inhibition rate,apoptosis rate,and miR-499a-5p expression of hemangioma endothelial cells were significantly increased in a dose-dependent manner(P<0.05).After overexpression of miR-499a-5p,the proliferation inhibition rate and apoptosis rate of hemangioma endothelial cells were significantly increased(P<0.05).After inhibition of miR-499a-5p,the effect of MSA on the proliferation and apoptosis of hemangio endothelial cells was partially reversed(P<0.05).Conclusion MSA could inhibit the proliferation and promote apoptosis of endothelial cells in infantile hemangiomas,the mechanism may be related to the up-regulation of miR-499a-5p expression.