- VernacularTitle:5-氟尿嘧啶对肝癌细胞系BEL-7402中肿瘤干细胞的富集作用
- Author:
Yue YANG
1
;
De-long LI
;
Wen-jing ZHU
;
Fei LIU
;
Meng-tian KANG
;
Sen ZHAO
;
Pu-chen HAO
;
Xu HAN
;
Pei-qiong CHEN
;
Fu-dong LÜ
;
Ji-liang FENG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Cycle; drug effects; Cell Line, Tumor; Cell Proliferation; drug effects; Fluorouracil; pharmacology; Humans; Neoplastic Stem Cells; cytology; drug effects; metabolism
- From: Chinese Journal of Hepatology 2011;19(9):686-691
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of 5-FU (5-fluorouracil) on enriching cancer stem cells of HCC cell line BEL-7402 and the biological characteristics of enriched cells.
METHODSThe enriching concentration of 5-FU was determined by CCK-8 (cell counting kit-8). Flow Cytometry was used to determine the changes in cell cycle and positive expression ratio of surface marker CD56, CD54, EpCAM and CD133. The self-renewal and differentiation of positive cells were tested by colony formation assay, and were compared with the control group.
RESULTSEnriching concentration of 5-FU was determined as 10 μg/ml with 48 h incubation. After enrichment, G0/G1 phase cells increased from 57.50 %+/-0.98% to 68.70%+/-3.41% (P<0.05). Whereas S phase cells decreased from 40.26%+/-4.12% to 31.80%+/-4.15% (P<0.01); G2/M phase cells disappeared in experimental group, and was 5.80%+/-1.87% in control group (P<0.01). The proportion of the cell cycle changed with significant statistical differences. Meanwhile, positive rate of cell surface makers CD56, CD54, EpCAM and CD133 increased from 0.57%+/-0.12%, 8.10%+/-6.79%, 0.3%+/-0.01% and 3.20%+/-0.99% to 4.13%+/-0.06%, 50.08%+/-1.69%, 0.55%+/-0.07% and 10.51%+/-1.13%, respectively. The difference was significant (P<0.05). The colony forming ratio of CD56, CD54, EpCAM and CD133 negative cells and positive cells were 2.11%+/-0.21%, 3.32%+/-0.31%; 0.86%+/-0.101%, 2.40%+/-0.52 %; 7.19%+/-0.56%, 7.73%+/-0.71%; 2.70%+/-0.26%, 5.75%+/-0.81%, respectively, and significant differences were found between (P<0.05).
CONCLUSION5-fluorouracil enriched the cancer stem cell population in HCC cell line BEL-7402. CD56 and CD54 can be used as important surface markers in research of liver cancer stem cells.