维生素D调控Wnt/β-catenin信号通路抑制乳腺癌干细胞活性  

Vitamin D inhibited breast cancer stem cells through Wnt/β-catenin pathway

作　　者：沙昱彤 黄家明 卢珍萍 梁欢 征宗梅 杨学攀 施洪飞[1] 朱明明 SHA Yutong;HUANG Jiaming;LU Zhenping;LIANG Huan;ZHENG Zongmei;YANG Xuepan;SHI Hongfei;ZHU Mingming(Department of Nutrition,the Second School of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学第二临床医学院营养教研室

出　　处：《中国癌症防治杂志》2019年第4期297-302,共6页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：国家自然科学基金项目(81803217);江苏省高等学校自然科学研究项目(18KJB330003)

摘　　要：目的探讨维生素D对乳腺癌干细胞活性的影响及其可能的作用机制。方法采用MTT法检测不同浓度(1 μmol/L、5 μmol/L、10 μmol/L和20 μmol/L)维生素D对乳腺癌SUM159细胞活力的影响,用无血清培养基悬浮培养富集乳腺癌干细胞CSCSUM159,分析维生素D对乳腺癌干细胞活性的影响。Western blot和Real-time qPCR 分别检测乳腺癌干细胞标志物CD133、CD44、Oct-4、Nanog以及Wnt/β-catenin信号通路相关分子p-GSK-3β、β-catenin、c-Myc和cyclin-D1的表达。结果 MTT法检测结果显示,1 μmol/L、5 μmol/L、10 μmol/L和20 μmol/L维生素D均可显著抑制乳腺癌SUM159细胞的活力(P<0.01)。无血清悬浮培养可有效富集乳腺癌干细胞CSCSUM159。维生素D可使乳腺癌干细胞CSCSUM159的成球数量减少,且细胞成球大小较对照组小。Western blot和Real-time qPCR检测结果显示,维生素D干预后,乳腺癌干细胞CSCSUM159中CD133、CD44和Nanog的蛋白表达下调,Wnt/β-catenin信号通路相关分子p-GSK-3β、β-catenin和c-Myc蛋白表达及β-catenin、c-Myc和cyclin-D1 mRNA表达亦下调(P<0.05)。结论维生素D可抑制乳腺癌干细胞活性,可能与其调控Wnt/β-catenin信号通路活性有关。Objective To explore the effect of Vitamin D on the activity of breast cancer stem cells and its possible mechanism. Methods The effects of different concentrations(1 μmol/L, 5 μmol/L, 10 μmol/L and 20 μmol/L) of Vitamin D on the viability of breast cancer SUM159 cells were detected by MTT assay. The breast cancer stem cells CSCSUM159 were enriched in suspension culture in serum-free medium to analyze the effect of Vitamin D on the activity of breast cancer stem cells. Western blot and Real-time qPCR were used to detect the expression of the breast cancer stem cell markers CD133, CD44, Oct-4 and Nanog,and Wnt/β-catenin signaling pathway-related genes p-GSK-3β,β-catenin, c-Myc and cyclin-D1. Results MTT assay showed that 1 μmol/L, 5 μmol/L, 10 μmol/L and 20 μmol/L Vitamin D inhibited the viability of breast cancer SUM159 cells significantly(P<0.01). Serum-free medium culture could effectively enrich breast cancer stem cells CSCSUM159. Vitamin D reduced the number and size of SUM159 tumorsphere. Western blot and Real-time qPCR showed that Vitamin D down-regulated the protein expression of CD133, CD44 and Nanog and inhibited the protein expression levels of p-GSK-3β,β-catenin and c-Myc,and down-regulated the mRNA expression of β-catenin,c-Myc and cyclin-D1 in breast cancer stem cell CSCSUM159(P<0.05). Conclusion Vitamin D can inhibit the activity of breast cancer stem cells, which may be related to its regulation of Wnt/β-catenin signaling pathway.

关 键 词：乳腺癌 维生素D 肿瘤干细胞 WNT/Β-CATENIN信号通路 

分 类 号：R737.9[医药卫生—肿瘤]