纳米金结合重组人血管内皮抑素对人脐静脉内皮细胞增殖、凋亡及迁移的影响  

The effect of the gold nanoparticles combined with endostar on proliferation, apoptosis, migrationof human umbilical vein endothelial cells

作　　者：刘书昊[1] 潘运龙[1] 覃莉[2] 李伟[1] 阳小燕 李鑫[1] 杨文德[1] 潘璠 Liu Shuhao;Pan Yunlong;Qin Li;Li Wei;Yang Xiaoyan;Li Xin;Yang Wende;Pan Fan(Department of Gastrointestinal Surgery, the First Affiliated Hospital of Jinan University, Guangzhou 510630, China;Department of Histology and Embry- ology, Medical School of Jinan University, Guangzhou 510632, China)

机构地区：[1]暨南大学附属第一医院胃肠外科,广州510630 [2]暨南大学医学院组织胚胎学教研室,广州510632

出　　处：《中华实验外科杂志》2018年第4期645-648,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81472849）;广东省自然科学基金（2014A030313383）;广东省高水平大学建设项目基金（88016013034）

摘　　要：目的观察纳米金结合重组人血管内皮抑素对原代人脐静脉血管内皮细胞（HUVECs）增殖、凋亡及迁移的影响，并探讨其机制。 方法将原代HUVECs培养传至第4代，分为对照组、重组人血管内皮抑素组（ES）、纳米金组（AU）和纳米金结合重组人血管内皮抑素组（AU-ES）。采用细胞计数试剂盒（CCK-8）法分别检测在24、48、72 h和25、50、100 μg/ml下各组细胞增殖抑制率，可得出最佳时间及浓度组合条件；在该条件下采用流式细胞仪、Transwell实验观察各组HUVECs的凋亡率、迁移能力的变化；采用Western blot检测各组与细胞增殖、凋亡及迁移功能相关蛋白质表达水平变化。 结果（1）CCK-8结果表明在不同浓度及时间下，AU-ES组（24 h，100 μg/ml）增殖抑制率为（35.166±3.076）%，差异有统计学意义（t＝4.251，P＝0.013）；（24 h，100 μg/ml）为最佳处理条件；（2）流式细胞仪结果表明在24 h，100 μg/ml下，AU-ES组对HUVECs的凋亡率为（27.197±0.390）%，与对照组[（10.357±3.012）%]比较，差异有统计学意义（t＝9.603，P＝0.001）；（3）Transwell实验24 h后小室底膜迁移面积比为（42.943±3.129）%，与对照组（58.98±1.543）%比较，差异有统计学意义（t＝7.963，P＝0.001）；（4）与细胞增殖、凋亡及迁移有关蛋白如：B细胞淋巴瘤/白血病-2（bcl-2）、c-Myc、基质金属蛋白酶-2（MMP-2）、血管内皮因子生长受体-2（VEGFR-2）、波形蛋白（Vimentin），与对照组比较AU-ES组的蛋白表达水平是明显降低。 结论在一定条件下AU-ES能对原代HUVECs具有抑制增殖及迁移、促进凋亡的作用，并且降低有关蛋白的表达。ObjectiveTo observe the effect of the gold nanoparticles combined with endostar on proliferation, apoptosis, migration of human umbilical vein endothelial cells （HUVECs）, discuss the mechanism. MethodsFirstly, we cultivated the primary human umbilical vein endothelial cells to the fourth generation. The group divided into endostar group （ES）, gold nanoparticles group （AU）, gold nanoparticles combined with endostar group （AU-ES） and blank control group. The proliferation rate of HUVECs was investigated by cell counting kit-8 （CCK-8） assay under different time and different concentration, the optimum treatment concentration and time were selected. Under the optimum treatment condition, the apoptosis rate and the cell migration ability were assessed by flow cytometer, the Transwell assay. The expression level of function related proteins were examined by Western blotting assay. Results（1）CCK-8 assay results showed that in different concentration and time, AU-ES group （24 h, 100 μg/ml） proliferation inhibition rate was （35.166±3.076）%, the difference was statistically significant （t=4.251, P=0.013）, and （24 h, 100 μg/ml） was the optimum condition; （2）Flow cytometry results showed that in （24 h, 100 g/ml）, the apoptosis rate of HUVECs in AU-ES group was （27.197±0.390）%, and compared with the control group （10.357±3.012）%, the difference was statistically significant （t=9.603, P=0.001）; （3）The Transwell experiment results showed the migration cell area ratio was （42.943±3.129）%, and compared with the control group （58.980±1.543）%, the difference was statistically significant （t=7.963, P=0.001）; （4）AU-ES could decrease the expression of proliferation, apoptosis and migration related proteins [B-cell lymphoma/leukemia-2 （bcl-2）, c-Myc, matrix metalloproteinase-2 （MMP-2）, vascular endothelial growth factor receptor-2 （VEGFR-2）, Vimentin]. ConclusionUnder certain condition, gold nanoparticles combined with endostar c

关 键 词：纳米金 重组人血管内皮抑素 血管内皮细胞 增殖 凋亡 迁移 

分 类 号：R73[医药卫生—肿瘤]