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目的探讨组蛋白去乙酰化酶抑制剂(Histone deacetylases inhibitor,HDACi)辛二酰苯胺氧肟酸(Suberoy-lanilide hydroxamic acid,SAHA)对人脐静脉内皮细胞(Human umbilical vein endothelial cell,HUVEC)增殖及血管形成能力的影响。方法收集处于对数生长期的HUVEC,以不同浓度的SAHA分别处理24和48 h,另设不加SAHA的对照组,采用CCK-8法检测细胞的增殖活力,并计算增殖抑制率和半数抑制浓度(IC50)。采用流式细胞术检测经15μmol/L SAHA处理48 h的HUVEC凋亡和细胞周期,基质胶体外血管生成试验检测HUVEC的体外成管能力,Western blot法检测HUVEC细胞周期及凋亡相关蛋白的表达水平。结果随着SAHA浓度的增加及作用时间的延长,其对HUVEC增殖的抑制作用增强,SAHA浓度高于80μmol/L时,抑制率增加不明显,24和48 h的IC50值分别为60.53和30.49μmol/L;经15μmol/L SAHA处理48 h,与对照组相比,HUVEC的凋亡率明显增加(P<0.001),S期细胞比例明显升高(P<0.001),G0/G1期比例明显降低(P<0.001),体外成管能力明显下降,P21、caspase-3激活型、caspase-9酶原和激活型蛋白的表达水平均明显升高(P<0.001)。结论 SAHA能够抑制HUVEC增殖及体外血管形成能力,并使P21、caspase-3和caspase-9蛋白水平上调,为肿瘤的治疗提供了一种新的思路。
Objective To investigate the effect of HDACi suberoy-lanilide hydroxamic acid (SAHA) on the proliferation of human umbilical vein endothelial cells (HUVEC) Effect of vascular formation ability. Methods HUVECs in logarithmic growth phase were collected and treated with different concentrations of SAHA for 24 and 48 h, respectively. A control group without SAHA was also established. The viability of cells was detected by CCK-8 assay and the proliferation inhibition rate and half inhibition Concentration (IC50). The apoptosis and cell cycle of HUVECs treated with 15μmol / L SAHA for 48 h were measured by flow cytometry. The in vitro tube formation ability of HUVECs was detected by using matrix colloid in vitro. Western blot was used to detect the cell cycle and the expression of apoptosis related proteins Level. Results With the increase of SAHA concentration and the prolongation of action time, the inhibitory effect on the proliferation of HUVECs was enhanced. When the concentration of SAHA was higher than 80μmol / L, the inhibition rate did not increase obviously. The IC50 values at 24 and 48 h were 60.53 and 30.49μmol / L; After treated with 15μmol / L SAHA for 48h, the apoptosis rate of HUVEC increased significantly (P <0.001), the proportion of S phase cells increased significantly (P <0.001) (P <0.001). The ability of tube formation in vitro was significantly decreased. The expressions of P21, caspase-3, caspase-9 and activation protein were significantly increased (P <0.001). Conclusions SAHA can inhibit HUVEC proliferation and angiogenesis in vitro and upregulate the levels of P21, caspase-3 and caspase-9, providing a new idea for the treatment of cancer.