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Isolation of side population cells and detection of ABCG2 from SW480

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Chinese Journal of Cancer Research

Abstract

Objective

Side population cells (SP cells) are a new type of stem cells. They mainly express ABCG2/BCRP1 and have the ability to eliminate DNA dye Hoechst33342. Many studies showed that side population cells were able of self-renewal, differentiation and carcinogenesis in cancers. Our investigation aimed at isolation of side population cells and ABCG2 positive subpopulation from colon cancer cell line SW480 and identification of their characteristics of cancer stem cells.

Methods

Side population cells and non-side population cells of colon cancer cell line SW480 were isolated with DNA dye Hoechst33342 and their cell cycles were measured by flow cytometry. Expression of ABCG2 of SW480 was measured by immunohistochemistry and immunofluorescence, and its proportion was measured by flow cytometry.

Results

SW480 contained 2.29% side population cells. The fraction of side population cells decreased greatly to 0.40% by treatment with verapamil. The fraction of side population cells in S-G2M cell cycle was 16.14%, which was much lower than the fraction (34.05%) of non-side population cells in S-G2M. In SW480, ABCG2 positive cells, which proportion was 9.66%, were small, circular or oval, lack of psuedopods, similar to poor differentiation. On the contrary, the ABCG2 negative cells were large, polygonal, with many psuedopods, similar to high differentiation.

Conclusion

Our assay identified that side population cells did exist in SW480 and had a quiescence characteristic of stem cells. ABCG2 positive subpopulation occupied about 9.66% of SW480 and may have the ability to promote cell self-renewal and inhibit cell differentiation. Therefore, to isolate ABCG2 positive subpopulation from side population cells may be an alternative to study colorectal cancer stem cells.

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Authors and Affiliations

  1. Department of Oncology, The First Affiliated Hospital, Wenzhou Medical College, Wenzhou, 325000, China

    Liu Hai-guang  (刘海光), Pan Yi-fei  (潘贻飞), Guo Gui-long  (郭贵龙), Hu Xiao-qu  (胡孝渠) & Zhang Xiao-hua  (张筱骅)

  2. Department of Pathology, The First Affiliated Hospital, Wenzhou Medical College, Wenzhou, 325000, China

    Huang Ka-te  (黄卡特)

Authors
  1. Liu Hai-guang  (刘海光)
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  2. Pan Yi-fei  (潘贻飞)
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  3. Guo Gui-long  (郭贵龙)
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  4. Hu Xiao-qu  (胡孝渠)
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  5. Huang Ka-te  (黄卡特)
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  6. Zhang Xiao-hua  (张筱骅)
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Correspondence to Zhang Xiao-hua  (张筱骅).

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Liu, Hg., Pan, Yf., Guo, Gl. et al. Isolation of side population cells and detection of ABCG2 from SW480. Chin. J. Cancer Res. 19, 238–243 (2007). https://doi.org/10.1007/s11670-007-0238-7

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  • DOI: https://doi.org/10.1007/s11670-007-0238-7

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