三黄煎剂通过下调Aurora激酶A抑制乳腺癌血管新生

Sanhuang Decoction Down-Regulates Aurora Kinase A to Inhibit Breast Cancer Angiogenesis

目的探索三黄煎剂抑制乳腺癌血管新生之功效,并探讨其内在机制。方法siRNA沉默乳腺癌细胞中Aurora激酶A,qRT-PCR、Western Blot实验分别检测三黄煎剂及Aurora激酶A敲减对乳腺癌细胞Aurora激酶A的mRNA、蛋白表达的影响。人脐静脉血管内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)迁移、管腔形成实验分别检测三黄煎剂及Aurora激酶A敲减对乳腺癌细胞条件培养基诱导下的HUVECs的迁移、管腔形成的影响。鸡胚尿囊膜(Chorioallantoic Membrane,CAM)模型检测三黄煎剂和/或Aurora激酶A敲减对乳腺癌血管新生的抑制效果。Western Blot实验检测三黄煎剂及Aurora激酶A敲减对细胞外调节蛋白激酶(Extracellular Regulated Protein Kinases,ERK)信号通路表达的影响。结果三黄煎剂能够显著抑制Aurora激酶A的mRNA、蛋白的表达,且效果与Aurora激酶A敲减近似。三黄煎剂能够抑制乳腺癌细胞条件培养基诱导下的HUVECs的迁移、管腔形成,且与Aurora基因敲减作用相似。三黄煎剂能够抑制CAM上乳腺癌血管新生,联合运用三黄煎剂和Aurora激酶A敲减并不显著增加抑制效果。三黄煎剂能够调节ERK信号通路表达,且与Aurora激酶A敲减效果类似。结论三黄煎剂可能是通过靶向性调节Aurora激酶A,下调ERK信号通路,从而抑制乳腺癌血管新生。

Objective To investigate the effect of Sanhuang Decoction(SHD)on inhibiting angiogenesis of breast cancer and its mechanism.Methods Aurora kinase A in breast cancer cells was knocked down by the siRNA,and qRT-PCR and Western Blot analysis were used to detect the expression of mRNA and protein of Aurora kinase A in breast cancer cells.Cell migration and Matrigel detection were used to determine the inhibition of the migration and tube formation of human umbilical vein endothelial cells(HUVECs).In addition,a chorioallantoic membrane(CAM)assay was carried out to directly reveal the effect of SHD on breast cancer anti-angiogenesis.Furthermore,the extracellular signal-regulated kinase(ERK)expression in breast cancer cells was examined by western blotting.Results SHD could significantly inhibit the expression of Aurora kinase A.SHD could inhibit the migration and tube formation of huma HUVECs induced by breast cancer cell condition medium,and the effect was similar to Aurora kinase A knockdown.Moreover,SHD treatment could inhibit tumor neovascularization in CAM model,and coadministration of SHD and si-RNA ARUKA could not notably improve the inhibition of angiogenesis in CAM model.Furthermore,SHD treatment mimicked the regulation of ERK expression by AURKA knockdown in breast cancer cells,which was examined by western blotting.Conclusion SHD may inhibit tumor angiogenesis in breast cancer by targeting Aurora kinase A and downregulating the ERK signaling pathway.