|Table of Contents|

Preliminary study on the accumulation and identification of breast cancer stem cells

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2019 06
Page:
929-934
Research Field:
Publishing date:

Info

Title:
Preliminary study on the accumulation and identification of breast cancer stem cells
Author(s):
Zhang Ying1; Dai Yinhai1; 2; Qiu Chunli1; 3; Wang Mao2; Ma Hailong2; Mao Ning2
1.Shaanxi University of Traditional Chinese Medicine，Shaanxi Xianyang 712000,China;2.The Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine,Shaanxi Xianyang 712000,China;3.Xianyang Central Hospital,Shaanxi Xianyang 712000,China.
Keywords:
breast cancer stem cells; flow cytometry; suspension culture; enrichment; identification
PACS:
R737.9
DOI:
10.3969/j.issn.1672-4992.2019.06.006
Abstract:
Objective:A method for culturing and enriching breast cancer stem cells was discovered by enriching and identifying breast cancer stem cells from MCF-7,ZR-75-1,and MDA-MB-231 breast cancer cell lines.Methods:MCF-7,ZR-75-1 and MDA-MB-231 cell lines were subjected to adherent culture,and the morphology of each cell was observed by an inverted microscope.CD44-CD24-,CD44-CD24+,CD44+CD24- and CD44+CD24+ cells were collected by flow cytometry,in which CD44+CD24- were breast cancer stem cells,and the other three groups were the control group.Cells were counted by MTT method,and the growth curve of MCF-7,ZR-75-1 and MDA-MB-231 cell lines was drawn.MCF-7 cell line was subjected to serum-free suspension culture for 1 cycle,and the content of molecular surface marker CD44+CD24- was detected by flow cytometry.The inherently cultured CD44+CD24-breast cancer stem cells were used as the control group.The sorted MCF-7 cells(CD44+CD24-) and the sorted remaining MCF-7 cells(the sorted remaining MCF-7 cells) were subjected to stemness sphere-forming assays to identify CD44+CD24-stemness expression.Results:MCF-7 and MDA-MB-231 cell lines were rich in breast cancer cells of surface marker CD44-CD24-.ZR-75-1 cell line were rich in breast cancer cells of surface marker CD44+CD24+.Growth curve showed that MCF-7 ZR-75-1 and MDA-MB-231 cells took on continuous increase,and MDA-MB-231 cells grew faster than MCF-7 and ZR-75-1 cells.CD44+CD24-breast cancer stem cells were enriched from 19.4% to 88.9% by serum-free suspension culture.The number of CD44+CD24- phenotype cells in the sphere-forming assay was significantly higher than that of the sorted remaining MCF-7 cells,with the sphere-forming rate of (36.5±1.7)%，（1.1±0.5）%.Conclusion:Flow cytometry can successfully sort out breast cancer stem cells with molecular surface marker CD44+CD24-.Different subtypes of breast cancer vary in gene expression,invasion,metastasis,hormone sensitivity and prognosis.CD44+CD24- may not be the only surface marker of breast cancer stem cells.MDA-MB-231 cell line grows faster than MCF-7,ZR-75-1 cell lines.Serum-free suspension culture method can enrich breast cancer stem cells easily and efficiently.CD44+CD24- breast cancer stem cells have stronger stemness expression.

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Memo

Memo:
陕西省教育厅专项科研计划项目（编号：17JK0209）;陕西省卫生科研项目（编号：2016D031）
Last Update: 1900-01-01