实用肿瘤学杂志 ›› 2012, Vol. 26 ›› Issue (5): 432-437.doi: 10.3969/j.issn.1002-3070.2012.05.011

• 论著 • 上一篇    下一篇

乳腺癌细胞系中肿瘤干细胞相关亚群的初步研究

陈炬莹, 陈晓耕   

  1. 福建医科大学附属龙岩市第一医院普外三科(龙岩山 364000)
  • 收稿日期:2011-09-05 出版日期:2012-05-25 发布日期:2012-10-29
  • 通讯作者: 陈炬莹,E-mail:chen-juy@163.com
  • 作者简介:陈炬莹,男,(1975-),硕士,主治医师,从事甲状腺乳腺治疗方面的研究

Preliminary study on cancer Stem-like cells in human breast cancer cell Iines

CHEN Juying,CHEN Xiaogeng   

  1. The First Affiliated Hospital of Longyan of Fujian Medical University,Longyan 364000,China
  • Received:2011-09-05 Online:2012-05-25 Published:2012-10-29

摘要: 目的 通过乳腺癌细胞系及其干细胞的培养,化疗药干预和流式细胞仪筛选鉴定,探讨不同乳腺癌细胞系中CD44+CD24-/low细胞比例,及富集乳腺癌干细胞相关亚群的方法。方法 通过细胞培养乳腺癌细胞系MCF-7、MDA-MB-231,观察生长曲线,比较化疗药物干预下生长情况;利用流式细胞仪检测两种乳腺癌细胞系中CD44+CD24-/low细胞比例;无血清悬浮培养,化疗药(多西紫杉醇、表阿霉素)干预这两种乳腺癌细胞系,观察其是否形成细胞球。结果 (1)MDA-MB-231细胞系倍增时间短,生长速率高于MCF-7细胞系;(2)MCF-7细胞系中可能存在较大比例肿瘤干细胞,其对化疗抵制,能自我更新;(3)化疗敏感性用两独立样本t检验,MCF-7细胞,差异没有统计学意义;MDA-MB-231细胞,差异有统计学意义,提示MDA-MB-231细胞系对该方案化疗较敏感;(4)无血清悬浮培养,MDA-MB-231细胞系未发现明显细胞球;MCF-7细胞系初次无血清培养约6天出现细胞球。加入化疗药筛选后两种细胞,见大部分肿瘤细胞逐渐死亡,未发现明显细胞球;(5)流式细胞仪检测,MCF-7、MDA-MB-231两种细胞系中主要是CD44+CD24+亚群和CD44-CD24+亚群,CD44+CD24-/low细胞比例分别2.07%和0.20%。结论 (1)MDA-MB-231细胞系增值较快,恶性度相对较高,其对TA联合化疗药物较敏感;MCF-7细胞系中可能存在少量肿瘤干细胞,对化疗抵制,能自我更新;(2)无血清培养液培养MCF-7细胞系能形成悬浮细胞球;流式细胞仪检测两种细胞系中CD44+CD24-/low细胞比例小;(3)CD44+CD24-/low表型可能不是乳腺癌干细胞唯一特异性的表面标志。

Abstract: Objective Under culturing different human breast cancer cell lines and the related content of cancer stem-like cells,we discuse CD44+CD24-/low cell proportion in breast cancer cell lines and related subsets enrichment method of breast cancer stem-like cells through effective intervention of the commonly chemotherapy and flow cytometric analysis.Methods Through the cell culture of breast cancer cell line“MCF - 7,MDA - MB -231”,we observed the growth curve and compared grow under conditions with chemotherapy drugs intervention.we detected the expression proportion of CD44+CD24-/low cell in the two breast cancer cell lines by flow cytometry.Under suspension culturing the above cell lines in serum-free medium,we observed clonal nonadherent spherical clusters by combining with or without chemotherapy scheme of TA scheme(docetaxel+table doxorubicin).Results 1.The multiplial time of MDA-MB-231 cells was shorter,and its growth rate was higher than MCF - 7 cells.2.The MCF-7 cells might exist the larger proportion of cancer stem-like cells with chemotherapy resistance and self-update.3.Chemotherapy sensitivity with two independent sample t-test,the result in MCF-7 cells suggested no statistical significance(P>0.05).The result in MDA-MB-231 cells showed obvious positive significance(P<0.05).MDA-MB-231 cells were more sensitive to chemotherapy from them.4.Under serum-free suspension cultures of two breast cell lines,MDA-MB-231 cells got no clear cell balls.MCF-7 cells were found cell balls at the first 6 day.With chemotherapy,the majority of the two tumor cells gradually died,and there were no living cell balls.5.Measured the content of CD44+CD24-/lowcytometry cells by flow cytometry,the two above cell lines mainly contain CD44+CD24+ subsets(74.2%,95.7%)and CD44-CD24+ subsets(23.3%,3.95%).The CD44+CD24-/low proportion of MCF-7 and MDA-MB-231 was 2.07%and 0.20%,respectirely.Conclusion 1.The MDA-MB-231 cells line which growed faster had higher malignant degree and showed more sensitive than MCF-7 cells line to TA chemotherapy scheme.2.Under serum-free suspension cultures,MCF-7 cells grow clonal monadherent spherical clusters,and the two lines had small proportion of CD44+CD24-/low cell with flow cytometric detection.3.The CD44+CD24-/low phenotype may not be the only specific surface markers of breast cancer stem cells.

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