肿瘤抑素影响婴幼儿血管瘤内皮细胞增殖与凋亡的相关机制

Effect of tumstatin on proliferation and apoptosis of endothelial cell in infantile hemangioma

目的观察肿瘤抑素对婴幼儿血管瘤内皮细胞增殖及凋亡的影响,并对相关机制进行初步探讨。方法采用0、5、10、20和40μg/ml的肿瘤抑素及40μg/ml的肿瘤抑素处理婴幼儿血管瘤内皮细胞0、12、24和48 h,采用MTT方法及流式细胞术检测婴幼儿血管瘤内皮细胞的增殖及凋亡情况。采用Western blot方法检测婴幼儿血管瘤内皮细胞中AMPK和m TOR的磷酸化水平。结果同0μg/ml肿瘤抑素处理组相比,5、10、20和40μg/ml的肿瘤抑素均可显著抑制婴幼儿血管瘤内皮细胞的增殖能力,并促进婴幼儿血管瘤内皮细胞的凋亡(P<0.05),且这种作用具有计量依赖性。同0 h处理组相比,40μg/ml的肿瘤抑素处理婴幼儿血管瘤内皮细胞2、24和48 h均可显著抑制婴幼儿血管瘤内皮细胞的增殖能力,并促进婴幼儿血管瘤内皮细胞的凋亡(P<0.05),且这种作用具有时间依赖性。同对照相比,肿瘤抑素可上调婴幼儿血管瘤内皮细胞中AMPK的磷酸化水平,下调m TOR的磷酸化水平。结论肿瘤抑素可通过激活AMPK通路,进而抑制m TOR通路,从而抑制婴幼儿血管瘤内皮细胞的增殖能力,并促进婴幼儿血管瘤内皮细胞的凋亡。

[ Objective ] To observe the effect of tumstatin on proliferation and apoptosis of endothelial cell in infantile hemangioma, and study the related mechanism. [Methods] Endothelial cell in infantile hemangioma was treated with tumstatin with different concentrations （0, 5, 10, 20 and 40 p.g/ml） or in a time dependent manner （0, 12, 24 and 48 h）, then the proliferation ability and apoptosis of endothelial cell in infantile hemangioma were detected by MTT and FCM, phosphorylation level of AMPK and roTOR were detected by Western blot. [Results] Compared with 0 μg/ml tumstatin treated group, 5, 10, 20 and 40 μg/ml tumstatin treated can inhibit proliferation in endothelial cell in infantile hemangioma, and promote apoptosis of endothelial cell in infantile hemangioma （P 〈 0.05）, and those effect were concentration dependent. Compared with 0 h tumstatin treated group, 12, 24 and 48 h tumstatin treated can inhibit proliferation in endothelial cell in infantile hemangioma, and promote apoptosis of endothelial cell in infantile hemangioma （P 〈 0.05）, and those effect were time dependent. Compared with control group, tumstatin treated can active AMPK signaling pathway and inhibit roTOR signaling pathway in endothelial cell in infantile hemangioma. [ Conclusion] By active AMPK signaling pathway and inhibit roTOR signaling pathway, tumstatin can inhibit proliferation in endothelial cell in infantile hemangioma, and promote apoptosis of endothelial cell in infantile hemangioma.