千金藤素对黑色素瘤干性的影响及机制研究

The Mechanism Research on Cell Stemness Induced by Cepharanthine in Melanoma

摘要: 探讨药物千金藤素对人皮肤黑色素瘤细胞的作用及机制。使用CCK8实验检测人皮肤黑色素瘤细胞系A375对千金藤素的剂量毒性效应，将加入等体积DMSO的A375细胞定义为对照组，A375细胞以梯度浓度和时间的千金藤素处理组设为实验组。成球实验检测肿瘤细胞干细胞成球能力；EdU实验与克隆形成实验检测细胞增殖；蛋白免疫印记实验检测干细胞标记物，以及下游的信号转导机制。结果显示A375细胞随着千金藤素处理剂量与时间的增多增殖受到明显抑制，干性降低，表现为千金藤素处理组EdU标记新生细胞比例降低，肿瘤球形成能力下降，蛋白印迹显示干性标记物表达水平降低。通过对信号转导研究发现，实验组中JAK2/STAT3信号轴被抑制，表明千金藤素可能通过靶向JAK2/STAT3信号轴在人皮肤黑色素瘤中发挥抗癌作用。

Abstract: To investigate the effect of Cepharanthine on human melanoma cells, CCK8 assay was used to detect the dose toxicity of Cepharanthine in human melanoma cell line A375. A375 cells treated with equal volume of DMSO were defined as the control group, and A375 cells treated with gradient concentration and time of Cepharanthine were set as the experimental group. Tumor sphere formation assay was used to detect the sphere formation ability of tumor cells. EdU assay and colony formation assay were used to detect cell proliferation. Western blot was used to detect stem cell markers and downstream signal transduction mechanisms. The results showed that the proliferation of A375 cells was significantly inhibited and the stemness of A375 cells was decreased with the increase of the dose and time of Cepharanthine treatment. The proportion of EdU-labeled cells and the ability of tumor sphere formation were decreased in the Cepharanthine treatment group. Signal transduction studies showed that the JAK2/STAT3 signaling axis was inhibited in the experimental group, indicating that Cepharanthine may play an anti-cancer role in melanoma by targeting the JAK2/STAT3 signaling axis.