Page 95 - 《中国药房》2022年4期

纳米雄黄对MCF-7乳腺癌干细胞的体外抑制作用

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邬 俊 1，2＊ ，王 贵 ，陈 毅 ，王唯婷 ，姜 爽 ，王晓波 （1.解放军联勤保障部队第967医院药剂科，辽宁
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大连 116021；2.大连医科大学药学院，辽宁 大连 116044）
中图分类号 R285.5；R965 文献标志码 A 文章编号 1001-0408（2022）04-0473-08
DOI 10.6039/j.issn.1001-0408.2022.04.15

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（1 mg/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
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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Western blot法检测乳腺癌干细
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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、80 mg/L 组
MCF-7亲本细胞及其干细胞（两者1 mg/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、10 mg/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
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细胞有所减少，其划痕愈合率、相对侵袭率（水飞雄黄 1 mg/L 组除外）、CD44 /CD24 细胞亚群比例均显著低于空白对照组（P＜
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0.01）；水飞雄黄、纳米雄黄2.5、10 mg/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
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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。
关键词 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
In vitro inhibitory effects of realgar nanoparticles on MCF-7 breast cancer stem cells
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WU Jun ，WANG Gui ，CHEN Yi ，WANG Weiting ，JIANG Shuang ，WANG Xiaobo（1. Dept.of Pharmacy，No.
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967 Hospital of Joint Logistics Support Force of the Chinese People’s Liberation Army，Liaoning Dalian
116021，China；2. School of Pharmacy，Dalian Medical University，Liaoning Dalian 116044，China）
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
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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 CD44 /CD24 subgroup were significantly lower than blank control group（P＜0.01）. The expressions
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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 nanoparticles can
significantly inhibit the proliferation of breast cancer stem
＊硕士研究生。研究方向：纳米药物抗肿瘤干细胞作用。E-mail：
wujun121619@163.com cells， the formulation and differential ability of mammo-
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#a 通信作者：副主任药师，博士。研究方向：抗肿瘤纳米制剂。 sphere，and reduce the proportion of CD44 /CD24 － subgroup.
电话：0411-39847132。E-mail：jshcrystal@qq.com The effect may be associated with the inhibition of migration
#b 通信作者：主任药师，博士生导师，博士。研究方向：抗肿瘤中 and invasion of breast cancer stem cells by inhibiting the
药制剂。电话：0411-39847000。E-mail：wxbbenson0653@sina.com expression of proteins related to epithelial mesenchymal

中国药房 2022年第33卷第4期 China Pharmacy 2022 Vol. 33 No. 4 ·473 ·