天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (6): 675-678.doi: 10.11958/20150221

• 细胞与分子生物学 • 上一篇    下一篇

盐霉素对乳腺癌 MCF-7 球囊细胞增殖与上皮间质转化作用的研究

马赫遥， 付婴子， 何苗， 颜媛媛， 江茜， 孙也之， 魏敏杰   

  1. 中国医科大学药学院药理教研室
  • 收稿日期:2015-10-29 修回日期:2016-01-12 出版日期:2016-06-15 发布日期:2016-07-04
  • 基金资助:
    国家自然科学基金资助项目 （81373427， 81572898）； 辽宁省高等学校优秀人才支持计划项目 （LJQ2014084）

The study on the role of salinomycin in the proliferation and epithelial-mesenchymal transition of breast cancer MCF-7 mammosphere cells

MA Heyao， FU Yingzi， HE Miao， YAN Yuanyuan， JIANG Qian， SUN Yezhi, WEI Minjie   

  1. Department of Pharmacology, School of Pharmacy, China Medical University, Shenyang 110122， China
  • Received:2015-10-29 Revised:2016-01-12 Published:2016-06-15 Online:2016-07-04

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摘要： 摘要：目的 探讨盐霉素（Salinomycin）对MCF-7球囊细胞（MCF-7 MS）的增殖与上皮间充质转化 (EMT）的作用。方法 通过乳腺癌MCF-7细胞无血清悬浮培养富集乳腺癌干细胞（BCSCs）得到MCF-7 MS；CCK-8 assay检测0、10、30、100、300、1 000、3 000及10 000 nmol/L的浓度盐霉素处理24 h对MCF-7 MS细胞生存率的影响，并计算半抑制浓度（IC50）值。 Western blot检测30 nmol/L、60 nmol/L终浓度的盐霉素对MCF-7 MS细胞上皮标志物E-钙黏蛋白(E-cadherin)和间质化标志物Snail的表达影响，以等体积的DMSO处理的MCF-7 MS细胞作为对照组。BALB/c裸鼠皮下接种MCF-7 MS细胞制备荷瘤鼠模型并分为对照组（等体积的生理盐水）和盐霉素给药组（5 mg/kg盐霉素），免疫组织化学染色检测瘤组织的Snail和E-cadherin的表达。结果 随着浓度的增加，MCF-7 MS细胞的存活率逐渐下降（P < 0.05），24 h的IC50值为989 n mol/L。与对照组比较，30和60 nmol/L的盐霉素均可增加E-cadherin的表达，而降低Snail的表达，且后者作用更明显（P < 0.05）。与对照组相比，盐霉素给药组荷瘤鼠瘤组织内的Snail的表达下调，而E-cadherin的表达升高（P < 0.01）。结论 盐霉素能够抑制具有BCSCs特性的MCF-7 MS细胞的增殖，且可抑制MCF-7 MS细胞的EMT，是靶向肿瘤干细胞潜在药物。

关键词: 乳腺肿瘤, 癌, 肿瘤干细胞, 细胞增殖, 上皮间充质转化, MCF-7 球囊细胞

Abstract: Abstract： Objective To investigate the effects of salinomycin on the cell proliferation and epithelial-mesenchymal transition (EMT) of MCF-7 mammosphere (MCF-7 MS). Methods Breast cancer MCF-7 cells were cultured in suspension in serum-free medium to obtain MCF-7 MS. The cell viability of MCF-7 MS cells treated with serial concentrations of 0, 10, 30,100, 300, 1 000, 3 000 and 10 000 nmol/L of salinomycin for 24 hours were detected by CCK-8 assay. The half maximal inhibitory concentration (IC50) was calculated. Western blot analysis was performed to detect the expression levels of E-cadherin and Snail in MCF-7 MS cells treated with 30 nmol/L and 60 nmol/L salinomycin. The same capacity of DMSO was added to MCF-7 MS as control group. The xenograft tumors from MCF-7 MS transplant mice were divided into control group (the same capacity of normal saline) and salinomycin group (5 mg/kg salinomycin), then the expressions of E-cadherin and Snail were dectected by immunohistochemical staining. Results With the increased concentration of salinomycin, the cell survival rate of MCF-7 MS cells decreased (P < 0.05). The IC50 after 24 h-treatment was 989 nmol/L. Both 30 and 60 nmol/L of salinomycin increased the expression of E-cadherin and decreased the expression of Snail compared with control group. In addition, 60 nmol/L treatment group showed more significant effect (P < 0.05). In xenograft tumors from MCF-7 MS transplant mice, the expression of Snail decreased, and E-cadherin increased in salinomycin treatment group compared with control group (P < 0.01). Conclusion Salinomycin can inhibit the cell proliferation and EMT in MCF-7 MS cells, which is a potential drug to target cancer stem cells.

Key words: breast neoplasms, caricinoma, neoplastic stem cells, proliferation, epithelial-mesenchymal transition, MCF- 7 mammosphere