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

Issue:

2024 15

Page:

2722-2729

Research Field:

Publishing date:

Info

Title:

Methodological study on the isolation and identification of human breast cancer stem cells based on CD44 and ALDH1

Author(s):

JIANG Qin¹; ZHANG Manli¹; CHEN Hongqing²; ZHANG Xufan²; LI Sheng¹

1.Laboratory Department of Mianyang Traditional Chinese Medicine Hospital (Mianyang Hospital of Chengdu University of Traditional Chinese Medicine),Sichuan Mianyang 621000,China;2.Department of Laboratory Medicine,Hospital of Chengdu University of Traditional Chinese Medicine,Sichuan Chengdu 610075,China.

Keywords:

breast cancer stem cells; flow sorting; culture; oncogenic analysis; identification

PACS:

R737.9

DOI:

10.3969/j.issn.1672-4992.2024.15.006

Abstract:

Objective:To isolate and identify breast cancer stem cells from breast cancer tissues by molecular experiments,cell experiments,and animal experiments from four types of breast adenocarcinoma tissue samples.Aiming to find a treatment method for the study of breast cancer stem cells and provide directions for the treatment of breast cancer.Methods:Primary breast cancer single-cell suspension was obtained by DocSense tissue single-cell suspension preparation system for cell culture.CD44+/CD24-/low group,ALDH1+group and non stem cell phenotype CD44-/ALDH1- group were selected by flow cytometry using CD44/CD24 and ALDH1 antibodies.The MTT experiment draws the growth curves of the CD44+/CD24-/low,ALDH1+group,and CD44-/ALDH1-group screened out for Luminal A type,Luminal B type,HER-2+type,and Basal-like type respectively.Finally,nude mouse tumorigenesis experiments were performed,pathological sections of tumor tissue were peeled off,and hematoxylin-eosin staining (HE staining) was used to verify the stemness of the sorted cells and that the tissue was breast cancer tissue.Results:This study employed tissue single-cell ablation technology,stem cell tumor microsphere formation experiments,and flow cytometry,successfully enriching and identifying eight breast cancer stem cells across four molecular subtypes.Through microsphere clonal formation assays and MTT assays,breast cancer stem cells marked with the CD44+/CD24-/low,ALDH1+ phenotype were found to have stronger invasiveness and proliferative abilities.Subcutaneous tumor implantation in nude mice followed by HE staining further demonstrated that breast cancer cells with the CD44+/CD24-/low,ALDH1+phenotype exhibit enhanced tumorigenic capacity and pronounced heterogeneity.Conclusion:The CD44+/CD24-/low and ALDH1+groups have stronger stem cell expression in breast cancer,which can be used as the research direction for identifying and treating breast cancer stem cells.

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Last Update: 2024-06-28