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目的研究姜黄素对人脉络膜黑色素瘤细胞株OCM-1血管形成拟态(vasculogenic mimicry,VM)及其PI3K信号通路的影响。方法三维培养人脉络膜黑色素瘤细胞株OCM-1,观察其形成VM的能力。观察梯度浓度(0μmol·L~(-1)、10μmol·L~(-1)、20μmol·L~(-1)、30μmol·L~(-1)、40μmol·L~(-1))的姜黄素对OCM-1细胞株VM形成的影响,采用免疫细胞化学检测参与VM形成的PI3K与EPHA2的表达。结果姜黄素在体外可以抑制OCM-1细胞株VM的形成,呈浓度依赖性。姜黄素对OCM-1细胞Eph A2和PI3K蛋白表达的影响:Eph A2和PI3K蛋白表达随姜黄素(10μmol·L~(-1)、20μmol·L~(-1)、30μmol·L~(-1)、40μmol·L~(-1))浓度的升高呈浓度依赖性降低。各组分别与0μmol·L~(-1)姜黄素组比较,差异均有统计学意义(均为P=0.000),其中30μmol·L~(-1)姜黄素组和40μmol·L~(-1)姜黄素组比较,差异无统计学意义(P=0.300、P=0.188)。结论姜黄素能明显抑制OCM-1细胞株VM的形成,其机制可能与下调EPHA2、PI3K蛋白的表达,从而抑制PI3K信号通路有关。
Objective To investigate the effect of curcumin on vasculogenic mimicry (VM) and its PI3K signaling pathway in human choroidal melanoma cell line OCM-1. Methods Three-dimensional human choroidal melanoma cell line OCM-1 was cultured and its ability to form VM was observed. The effects of different concentrations (0μmol·L -1, 10μmol·L -1, 20μmol·L -1, 30μmol·L -1, 40μmol·L -1) The effect of curcumin on VM formation in OCM-1 cell line was examined by immunocytochemistry. The expression of PI3K and EPHA2 involved in VM formation was detected by immunocytochemistry. Results Curcumin inhibited the formation of VM in OCM-1 cells in vitro in a concentration-dependent manner. Effects of Curcumin on Eph A2 and PI3K Protein Expression in OCM-1 Cells: The expressions of Eph A2 and PI3K proteins were down-regulated with the increasing of curcumin (10 μmol·L -1, 20 μmol·L -1, 30 μmol·L -1) 1), 40μmol·L -1) concentration in a concentration-dependent manner. Compared with 0μmol·L -1 curcumin group, the differences were statistically significant (all P = 0.000), of which 30μmol·L -1 curcumin group and 40μmol·L -1 curcumin group, 1) curcumin group, the difference was not statistically significant (P = 0.300, P = 0.188). Conclusion Curcumin can significantly inhibit the VM formation in OCM-1 cells. The mechanism may be related to the down-regulation of the expression of EPHA2 and PI3K proteins and the inhibition of the PI3K signaling pathway.