三种抗肿瘤药物对MCF-7乳腺癌干细胞作用的初步研究  

Preliminary Study on the Effect of Three Kinds of Antineoplastic on MCF-7 Breast Cancer Stem Cells

作　　者：邬俊 陈毅 王贵 李双 姜爽 王晓波[2] WU Jun;CHEN Yi;WANG Gui;LI Shuang;JIANG Shuang;WANG Xiao-bo(School of Pharmacy,Dalian Medical University,Dalian 116044,China;Department of Pharmacy,the 967th Hospital of PLA Joint Logistics Support Force,Dalian 116021,China)

机构地区：[1]大连医科大学药学院,辽宁大连116044 [2]中国人民解放军联勤保障部队第967医院药剂科,辽宁大连116021

出　　处：《现代中药研究与实践》2022年第3期30-37,共8页Research and Practice on Chinese Medicines

摘　　要：目的通过无血清培养法富集MCF-7细胞系中乳腺癌干细胞(BCSCs),并考察阿霉素、5-氟脲嘧啶及纳米雄黄对其体外作用。方法以亲本细胞为对照,荧光显微镜以及流式细胞仪检测CD44及CD24表达、迁移及Transwell实验检测BCSCs迁移及侵袭能力;分化实验检测BCSCs分化和悬浮球形成能力;流式实验检测悬浮球培养、分化以及三种抗肿瘤药物作用后CD44^(+)/CD24^(-)比例变化;CCK-8法检测亲本、分化细胞及BCSCs增殖能力及三种抗肿瘤药物对其作用。结果亲本细胞主要为CD44^(+)/CD24^(+)亚群,悬浮球干细胞主要为CD44^(+)/CD24^(-)亚群;血清能诱导BCSCs进行分化使其呈贴壁生长,并降低CD44^(+)/CD24^(-)比例;与亲本细胞相比,BCSCs具有更强的生长增殖、迁移侵袭及耐药能力,三种抗肿瘤药中阿霉素对细胞增殖抑制作用最强,而10μg/mL纳米雄黄组降低CD44^(+)/CD24^(-)比例的作用更加突出。结论无血清培养法可有效富集乳腺癌干细胞,与亲本细胞相比,其具有更强的生长增殖、迁移侵袭及耐药能力,纳米雄黄可能是有效抑制BCSCs的药物之一。Objective To enrich breast cancer stem cells(BCSCs)from MCF-7 cell line by serum-free culture and investigate the effects of adriamycin,5-fluorouracil and realgar nanoparticles in vitro.Methods Using MCF-7 parental cells as control,the expression of CD44 and CD24 are detected by fluorescence microscope and flow cytometry,and the ability of migration and invasion of BCSCs are detected by migration test and Transwell invasion test;the ability of differentiation and mammosphere formation of BCSCs are detected by differentiation experiment;the changes of the proportion of CD44^(+)/CD24^(-)cell subsets after mammosphere culture,differentiation and the action of three antineoplastic are detected by flow cytometry;the proliferative ability of parents,mammosphere differentiated cells and BCSCs and the effects of three antineoplastic on them are detected by CCK-8 method.Results The parent cells are mainly CD44^(+)/CD24+subsets,and the mammosphere cells are mainly CD44^(+)/CD24^(-)subsets;Serum can induce the differentiation of BCSCs,make them become adherent cells,and reduce the ratio of CD44^(+)/CD24^(-)subsets;compared with parental cells,BCSCs have stronger growth,proliferation,migration,invasion and drug resistance.Among the three antineoplastic,adriamycin has the strongest inhibitory effect on the ability of proliferation of cells,while 10μg/mL nano realgar group has a more prominent effect on reducing the ratio of CD44^(+)/CD24^(-)subsets.Conclusion The serum-free culture method can effectively enrich breast cancer stem cells.Compared with the parental cells,it has stronger growth,proliferation,migration,invasion and drug resistance ability.Nano realgar may be one of the effective drugs to inhibit BCSCs.

关 键 词：乳腺癌干细胞 MCF-7细胞 无血清培养 纳米雄黄 

分 类 号：R285.5[医药卫生—中药学]