Initial identification of CD133+/CD44+ prostate cancer stem cell through magnetic bead cell sorting
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Initial identification of CD133+/CD44+ prostate cancer stem cell through magnetic bead cell sorting
Journal of Chongqing Medical UniversityVol. 36, Issue 5, Pages: 454-345(2013)
作者机构:
重庆医科大学附属第一医院泌尿外科,重庆,400016
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Published:2013,
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SHENG Xia, Wang Delin, LI Wenbin, et al. Initial identification of CD133+/CD44+ prostate cancer stem cell through magnetic bead cell sorting. [J]. Journal of Chongqing Medical University 36(5):454-345(2013)
DOI:
SHENG Xia, Wang Delin, LI Wenbin, et al. Initial identification of CD133+/CD44+ prostate cancer stem cell through magnetic bead cell sorting. [J]. Journal of Chongqing Medical University 36(5):454-345(2013)DOI:
Initial identification of CD133+/CD44+ prostate cancer stem cell through magnetic bead cell sorting
Objective:To sort the CD133+/CD44+ stem cell through magnetic bead cell sorting(MACS) from human prostate cancer cell line PC-3 and to lay the foundation for the further functional research. Methods:Expressions of CD133 and CD44 on the membrane of PC-3 cell before and after MACS were detected through flow cytometry(FCM). Formation of spheres cultured by serum-free medi-um(SFM) was observed and subsequent immunofluorescence(IF) assay was made. Differences in the morphology and proliferation capacity of PC-3 cell before and after MACS were distinguished. Expressions of prostatic acid phosphatase(PAP) before and after the differentiation assay were detected by immunohistochemistry(IHC) and Western blot. Results:Positive expressions of CD133 and CD44 of PC-3 cell were (1.33±0.05)% and (0.87±0.06)% through FCM,while those after MACS were (84.82±0.07)% and (99.91±0.03)% respectively. Positive expressions of CD133 and CD44 remained after SFM detected by IF. Proliferation capacity was higher in CD133+/CD44+ cell than in PC-3 cell(t=11,P=0.008) and CD133+/CD44+ cell after inducement(t=40.1,P=0.001). Ex-pressions of PAP of CD133+/CD44+ cell were positive after being induced by transforming growth factor-β and were negative in the uninduced CD133+/CD44+ cell based on IHC and Western blot. Conclusions:CD133+/CD44+ cell had some characteristics of stem cell after MACS from PC-3 cell line through initial functional identification,which could be matting for further explore of the prostate cancer stem cell.