姜黄素抗血管生成和黑色素瘤生长的分子机制探讨  

Effect and Mechanism of Curcumin on Melanoma Growth and Angiogenesis

作　　者：钦建伟[1] 陈琳[1] 

机构地区：[1]苏州大学附属第一医院,江苏苏州215006

出　　处：《中国实验方剂学杂志》2015年第9期118-123,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江苏省卫生厅医学科研项目(Z201304)

摘　　要：目的：探讨姜黄素（curcumin）对人脐静脉内皮细胞（HUVECs）增殖、迁移和血管生成以及黑色素瘤B16F10生长的影响。方法：以HUVECs细胞,B16F10细胞和B16F10鸡胚尿囊膜移植瘤组织为研究对象,用不同浓度姜黄素分别作用HUVECs细胞,B16F10细胞和B16F10鸡胚尿囊膜移植瘤组织,MTT法检测姜黄素对HUVECs的增殖抑制率;Hoechst 33258染色法观察HUVECs细胞核的形态学变化;Transwell小室迁移实验考察姜黄素对HUVECs迁移能力的影响;鸡胚尿囊膜（CAM）实验考察姜黄素对体内血管生成的抑制作用;采用Western blot法检测姜黄素对HUVECs细胞血管内皮细胞生长因子受体2（VEGFR-2）蛋白和B16F10细胞基质金属蛋白酶-2（MMP-2）蛋白的表达;采用B16F10鸡胚尿囊膜移植瘤模型分析姜黄素对肿瘤生长的影响,采用免疫组化和Western blot分析姜黄素对瘤组织内VEGFR-2,MMP-2蛋白表达的作用。结果：姜黄素（4~15μmol·L-1）对HUVECs增殖无抑制作用,但姜黄素8,10,15μmol·L-1能抑制VEGF诱导的HUVECs增殖（P〈0.05,P〈0.01）;Hoechst 33258染色结果显示姜黄素抑制VEGF诱导的HUVECs增殖不依赖于诱导HUVECs凋亡;Transwell迁移实验显示姜黄素（8,10,15μmol·L-1）能抑制HUVECs迁移（P〈0.05,P〈0.01）;姜黄素能减少血管数目,抑制体内血管生成,并且抑制HUVECs细胞VEGFR-2蛋白和B1610细胞MMP-2蛋白表达,与空白组比较均具有统计学差异（P〈0.05,P〈0.01）。姜黄素（20 mg·L^-1）能抑制黑色素瘤B16F10体内生长,免疫组化和Western blot结果显示姜黄素（20 mg·L^-1）能抑制瘤组织VEGFR-2和MMP-2蛋白表达（P〈0.01）。结论：姜黄素能抑制VEGF诱导的HUVECs增殖、迁移及血管生成,抑制黑色素瘤生长,其机制为抑制VEGFR-2,MMP-2蛋白表达。Objective： To investigate the effect of curcumin on the proliferation, migration, angiogenesis of human umbilical vein endothelial cells （HUVECs）, and the growth of B16F10 melanoma. Method： HUVECs were treatment with different concentrations （0-15 μmol· L^-1） of eurcumin and the proliferation was measured by MTT. The morphological change of apoptotie HUVECs was detecte by using Hoechst 33258. The effect of eurcumin on HUVECs migration ability was assayed by Transwell. The anti-angiogenesis action of curcumin was investigated by using the chick chorioallantoic membrane （CAM） assay. The expressions of vascular endothelial growth factor receptor 2 （VEGFR-2） in HUVECs and matrix metalloproteinase-2 （MMP-2） in B16F10 were measured by Western blot. B16F10 cells were inoculated into CAM and post tumor formation, the impact of curcumin on tumor growth was analyzed. The expressions of VEGFR-2 and MMP-2 in B16F10 tumor tissue were analyzed by immunohistoehemistry and Western blot. Result： Cureumin （4-15 μmol·L^-1） had no inhibitory effect on the proliferation of HUVECs, but it （8, 10, 15 μmol·L^-1） could inhibit VEGF-induced proliferation of HUVECs （P 〈 0.05, P 〈 0.01 ）. Hoechst 33258 staining showed that cureumin inhibited VEGF- induced the HUVECs, which was not dependent on the induction of HUVECs apoptosis （ P 〈 0.05, P 〈 0.01 ）. Transwell migration experiments showed that eureumin （8, 10, 15 μmol·L^-1） could inhibit HUVECs migration, CAM assay confirmed that curcumin could reduce the number of blood vessels, inhibited angiogenesis in vivo （P 〈0.05, P 〈0.01 ）. Curcumin could inhibit HUVECs VEGFR-2 and B1610 cells MMP-2 protein expressions （P 〈 0.05, P 〈0.01）. Curcumin （ 20 mg·L^-1） inhibited the growth of melanoma B16F10 in vivo, immunohistochemistry and Western blot showed that curcumin （20 mg·L^-1） inhibited the VEGFR-2 and MMP-2 protein expressions （P 〈 0.05, P 〈 0.01 ）. Conclusion： Curcumin inhibited VEGF-induced HUVECs

关 键 词：姜黄素 人脐静脉内皮细胞 血管生成 B16F10 

分 类 号：R285.5[医药卫生—中药学]