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纳米雄黄对MCF-7乳腺癌干细胞的体外抑制作用
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| 篇名： | 纳米雄黄对MCF-7乳腺癌干细胞的体外抑制作用 |
| TITLE： | In vitro inhibitory effects of realgar nanoparticles on MCF- 7 breast cancer stem cells |
| 摘要： | 目的 研究纳米雄黄对乳腺癌干细胞的体外抑制作用。方法以人乳腺癌MCF-7亲本细胞为对象，采用无血清培养法培养获得乳腺癌干细胞。以阿霉素（1mg/L）为阳性对照，以相同质量浓度的水飞雄黄为参照，采用CCK-8法检测纳米雄黄对MCF-7亲本细胞及干细胞增殖的影响；借助悬浮球形成及分化实验、划痕实验、Transwell侵袭实验及流式细胞仪检测纳米雄黄对乳腺癌干细胞悬浮球形成及分化能力、迁移及侵袭能力、CD44+/CD24－细胞亚群比例的影响；采用Westernblot法检测乳腺癌干细胞上皮间质转化通路相关蛋白（上皮钙黏着蛋白、波形蛋白）的表达水平。结果水飞雄黄、纳米雄黄1、5、10、40、60、80mg/L组MCF-7亲本细胞及其干细胞（两者1mg/mL组乳腺癌干细胞除外）的存活率均显著低于空白对照组（P＜0.01）；水飞雄黄、纳米雄黄1、2.5、5、10mg/L组的多细胞悬浮球（＞20个细胞）数量均显著少于空白对照组（P＜0.01），且悬浮球体积有所缩小，分化的贴壁细胞有所减少，其划痕愈合率、相对侵袭率（水飞雄黄1mg/L组除外）、CD44+/CD24－细胞亚群比例均显著低于空白对照组（P＜0.01）；水飞雄黄、纳米雄黄2.5、10mg/L组细胞中上皮钙黏着蛋白的表达水平均显著高于空白对照组，波形蛋白的表达水平均显著低于空白对照组（P＜0.01）。纳米雄黄的上述作用普遍优于相同质量浓度的水飞雄黄（P＜0.01）。结论与相同质量浓度的水飞雄黄比较，纳米雄黄可更明显地抑制乳腺癌干细胞的增殖、悬浮球的形成及分化能力，并可降低CD44+/CD24－细胞亚群比例。这种作用可能与纳米雄黄通过抑制上皮间质转化通路相关蛋白的表达而抑制乳腺癌干细胞的迁移和侵袭有关。 |
| ABSTRACT： | OBJECTIVE To study in vitro inhibitory effects of realgar nanoparticles on breast cancer stem cells. METHODS Human breast cancer MCF- 7 parent cells were selected as subjects and cultured by serum-free culture to obtain breast cancer stem cells. Using adriamycin （1 mg/L）as positive control ，same concentration of water-processed realgar as reference ，the effects of realgar nanoparticles on the proliferation of MCF- 7 parent cells and stem cells were detected by CCK- 8 method. The effects of realgar nanoparticles on the formation of mammosphere ，the ability of differentiation ，migration and invasion ，the proportion of CD44+/CD24－ subgroup in breast cancer stem cells were detected by mammosphere formation and differentiation experiment ， scratch experiment ，Transwell invasion experiment and flow cytometry. Western blot assay was used to detect the expression of proteins related to epithelial mesenchymal transformation pathway （E-cadherin and vimentin ） in breast cancer stem cells. RESULTS The survival rates of MCF- 7 parent cells and stem cells （except for breast cancer stem cells in both 1 mg/mL groups ）in 1，5，10，40，60，80 mg/L groups of water-processed realgar and realgar nanoparticles were significantly lower than blank control group（P＜0.01）. The number of mammosphere （＞20 stem cells ）in 1，2.5，5，10 mg/L groups of water-processed realgar and realgar nanoparticles was significantly lower than blank control group （P＜0.01）；the volume of mammosphere decreased and the differentiated adherent cells decreased ；the healing rate of wound ，relative invasion rate （except for water-processed realgar 1 mg/L group）and the proportion of CD 44+/CD24－ subgroup were significantly lower than blank control group （P＜0.01）. The expressions of E-cadherin in 2.5，10 mg/L groups of water-processed realgar and realgar nanoparticles was significantly higher than blank control group ，and the expressions of vimentin was significantly lower than those in blank control group （P＜0.01）. The above effects of realgar nanoparticles were generally better than those of water-processed realgar with the same mass concentration （P＜ 0.01）. CONCLUSIONS Compared with water-processed realgar with the same mass concentration ，realgar nanoparti cles can significantly inhibit the proliferation of breast cancer stem cells， the formulation and differential ability of mammo- sphere，and reduce the proportion of CD 44+/CD24－ subgroup. The effect may be associated with the inhibition of migration and invasion of breast cancer stem cells by inhibiting the expression of proteins related to epithelial mesenchymal transformation pathway. |
| 期刊： | 2022年第33卷第04期 |
| 作者： | 邬俊，王贵，陈毅，王唯婷，姜爽，王晓波 |
| AUTHORS： | WU Jun，WANG Gui，CHEN Yi，WANG Weiting ，JIANG Shuang ，WANG Xiaobo |
| 关键字： | 雄黄；纳米雄黄；水飞雄黄；乳腺癌干细胞；人乳腺癌MCF-7亲本细胞；侵袭；转移 |
| KEYWORDS： | realgar；realgar nanoparticles ；water-processed realgar ；breast cancerstem cell ；human breast cancer MCF- 7 parent |
| 阅读数： | 198 次 |
| 本月下载数： | 4 次 |
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