芹菜素对A375黑色素瘤生长、迁移和血管生成的影响及其作用机制

Effects of apigenin on growth, migration and angiogenesis of melanoma A375 cells and the mechanism

摘要:

目的  探讨芹菜素对A375黑色素瘤增殖、迁移和血管生成的影响及其作用机制。方法  采用四唑氮化合物（MTS）法检测芹菜素对人黑色素瘤A375细胞增殖的影响；裸鼠皮下移植瘤模型考察芹菜素对体内黑色素瘤生长的影响；划痕实验检测芹菜素对细胞体外迁移的影响；小管形成实验检测芹菜素对人黑色素瘤血管生成的影响；Western blot检测芹菜素对A375细胞中调控增殖及血管生成相关蛋白表达的影响，如血管内皮生长因子（VEGF）、基质金属蛋白酶-9（MMP-9）、磷酸化P38（p-P38）、总蛋白P38（t-P38）、磷酸化Erk（p-Erk）、总蛋白Erk（t-Erk）。结果  体外芹菜素处理A375细胞后，肿瘤细胞的体外增殖及体内生长较空白对照组明显受抑制；肿瘤细胞的迁移能力较溶剂对照组明显减弱；小管形成实验中，芹菜素能够抑制肿瘤细胞诱导的血管生成；分子机制研究发现，芹菜素能抑制血管生成相关蛋白VEGF及MMP-9的表达，同时芹菜素也能抑制调控肿瘤细胞增殖相关蛋白，如p-P38及p-Erk的表达。结论  芹菜素能抑制A375黑色素瘤生长、迁移及血管生成，其机制可能与抑制调控肿瘤细胞增殖及血管生成相关蛋白的表达有关。

Abstract:

Objective To study the effects of apigenin on proliferation, migration and angiogenesis of human melanoma A375 cells and the mechanism of action. Methods The effects of apigenin on the proliferation of A375 cells were evaluated by the MTS method, and those on the in vivo growth of melanoma were assessed by establishing a subcutaneous xenograft model of nude mice. The influence of apigenin on in vitro cell migration was detected using the scratch assay, and that on the angiogenesis of human melanoma was detected by the tube formation assay. The regulatory effects of apigenin on the expressions of proliferation- and angiogenesis-related proteins such as vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), phosphorylated p38 (p-p38), t-p38, phosphorylated ERK (p-ERK) and t-ERK were detected by Western blot. Results After in vitro treatment of A375 cells with apigenin, their in vitro proliferation and in vivo growth were significantly inhibited compared with those of the control group, and the migration capacity was also significantly lowered. Apigenin inhibited tumor cell-induced angiogenesis in the tube formation assay. Molecular mechanism study showed that apigenin inhibited the protein expressions of angiogenesis-related VEGF and MMP-9 as well as proliferation-related p-p38 and p-ERK. Conclusions Apigenin can inhibit the growth, migration and angiogenesis of melanoma A375 cells, probably by suppressing the expressions of proliferation- and angiogenesis-related proteins.