Sonic Hedgehog信号通路对小鼠血管瘤内皮细胞增殖的影响

Effect of Sonic Hedgehog signaling pathway on the proliferation of mouse hemangioma endothelial cells

目的探讨Sonic Hedgehog信号通路对小鼠血管瘤内皮细胞(EOMA细胞)增殖的影响。方法常规培养EOMA细胞,采用噻唑蓝(MTT)比色法检测Sonic Hedgehog信号通路抑制剂GANT61对EOMA细胞增殖的影响。采用EdU染色流式细胞术检测GANT61及Sonic Hedgehog信号通路激动剂SAG对EOMA细胞增殖的影响。采用实时荧光定量PCR法(RT-qPCR)检测GANT61及SAG对EOMA细胞增殖细胞核抗原(PCNA)及细胞周期蛋白D1(Cyclin D1)基因mRNA表达的影响。结果GANT61作用EOMA细胞后,细胞增殖受到抑制,且呈剂量及时间依赖性,24 h半数抑制浓度(IC50)为79.42μmol/L。EdU染色流式细胞术检测结果显示,GANT61干预可抑制EOMA细胞增殖,而SAG干预可促进EOMA细胞增殖。RT-qPCR检测结果显示,GANT61干预可抑制PCNA及Cyclin D1 mRNA表达,而SAG干预可促进PCNA及Cyclin D1 mRNA表达。结论Sonic Hedgehog信号通路可调控EOMA细胞增殖,其可能参与血管瘤的发生发展。

Objective To investigate the effect of Sonic Hedgehog signaling pathway on the proliferation of mouse hemangioma endothelial cells(EOMA cells).Methods EOMA cells were cultured regularly.Methyl thiazolyl tetrazolium(MTT)colorimetry was used to detect the effect of GANT61,a Sonic Hedgehog signaling pathway inhibitor,on the proliferation of EOMA cells.EdU staining and flow cytometry were used to detect the effects of GANT61 and Sonic Hedgehog signaling pathway agonist SAG on the proliferation of EOMA cells.The effects of GANT61 and SAG on the mRNA expressions of proliferating cell nuclear antigen(PCNA)and Cyclin D1 in EOMA cells were detected by real-time quantitative polymerase chain reaction(RT-qPCR).Results After EOMA cells were treated with GANT61,the proliferation of EOMA cells was inhibited in a dose-and time-dependent manner,and the 50%inhibitory concentration(IC50)value was 79.42μmol/L at 24 h.The detection results of EdU staining and flow cytometry showed that GANT61 intervention inhibited the proliferation of EOMA cells,and SAG intervention promoted the proliferation of EOMA cells.The detection results of RT-qPCR showed that GANT61 intervention inhibited the expressions of PCNA and Cyclin D1 mRNA,and SAG intervention promoted the expressions of PCNA and Cyclin D1 mRNA.Conclusion Sonic Hedgehog signaling pathway can regulate the proliferation of EOMA cells,which may be involved in the occurrence and development of hemangioma.