氧化亚铜纳米粒对B16细胞上皮间质转化的影响

Cuprous oxide nanoparticles effect on the epithelial-mesenchymal transition of B16 cells

目的制备氧化亚铜纳米粒(cuprous oxide nanoparticles,CONPs)并研究对小鼠黑色素瘤B16细胞上皮间质转化的影响。方法采用水热法制备氧化亚铜纳米粒。将B16细胞分为正常培养组和CONPs(5、25、50μg/ml)刺激培养组,于倒置相差显微镜下观察细胞形态学的变化,通过细胞划痕实验和Transwell检测CONPs对B16细胞迁移能力的影响,采用免疫荧光染色法和蛋白质印迹法检测B16细胞表型的相关分子标志物表达的变化。结果合成的氧化亚铜纳米粒分布均匀,粒径约40nm;体外细胞实验发现CONPs明显地抑制了B16细胞的迁移和侵袭能力;并刺激B16细胞上皮细胞表型的E-cadherin、Cytokeratin、Desmoplakin的表达明显升高,而间质细胞表型的N-cadherin、Vimentin的表达降低。结论 CONPs能明显地抑制黑色素瘤细胞的侵袭转移和上皮-间质转化(EMT)过程。

Objective To research the epithelial-mesenchymal transition （EMT） effects of cuprous oxide nanoparticles （CONPs） on melanoma.Methods Cuprous oxide nanoparticles were prepared hydrothermally.The B16 cells were cultured with cuprous oxide nanoparticlesat different concentrations （5,25,50 μg/ml）.The changes of the morphology of the B16 cell were observed under the inverted microscope.The effects of CONPs on B16 cell migration ability were detected through the Wound healing assay and the Transwell assay.Then cell immunofluorescence and western blotting were used to test the EMT related molecular markers, including E-cadherin, N-cadherin, Cytokeratin, and Vimentin.Results The synthesized cuprous oxide nanoparticles distribute uniformly with a diameter of 40 nm.Our study indicated that CONPs inhibited the EMT of B16 cell.A conversion process was discovered in this study.In B16 cells, CONPs inhibited B16 cell migration, promoted the expression of E-cadherin, Cytokeratin and Desmoplakin, while the expression of N-cadherin and Vimentin was repressed in protein level.Conclusion Cuprous oxide nanoparticles can significantly restrain the invasion and metastasis of melanoma cells and inhabit the EMT of B16 cells.