Expression of miR-590-3p in pancreatic cancer stem cells
10.3760/cma.j.issn.1674-1935.2011.04.006
- VernacularTitle:MiR-590-3p在胰腺癌干细胞中的表达
- Author:
Weiqiang GONG
;
Renyi QIN
;
Min WANG
;
Pui TIAN
;
Feng ZHU
;
Chengjian SHI
;
Zhifa ZHANG
;
Xu LI
;
Xiaoquan HONG
- Publication Type:Journal Article
- Keywords:
Pancreatic neoplasms;
Tumor stem cells;
MicroRNAs;
Cell spheres;
Serum-free medium
- From:
Chinese Journal of Pancreatology
2011;11(4):247-250
- CountryChina
- Language:Chinese
-
Abstract:
Objectives To isolate cancer stem cells (CSCs) in pancreatic cancer cell lines PANC1 and ASPC-1 with serum-free medium( SFM ), and to detect the expression of miR-590-3p in CSCs. Methods PANC1 and ASPC-1 cells was cultured in serum-free medium. The monoclonal formation, differentiation and cell cycle, half inhibitory concentration ( IC50 ), and the expression of the surface markers CD24 + , CD44 + were detected. qRT-PCR was used to detect the expression of miR-590-3p. Results After SFM culture, (0.94 ±0.53 ) % of ASPC-1 and (0.57 + 0. 12 ) % PANC1 survived, and they formed spheres, and could continuously passage in vitro. Cell spheres differentiation recurred when serum was supplemented in SFM. The G0/G1 stage proportion, CD24+ , CD44 + , CD24+ CD44+ cells proportion, IC50 in ASPC-1 cell were (75.3 ± 5.4)%,0.96% ~ 2.01%, 27.52% ~ 34.47%, 0.35% ~ 0.44% and (224.37 ± 5.71 ) μg/ml, which were significantly higher than that those in parent cell [ (43.7 ± 3.8 ) %, 0. 38% ~ 0.42%, 17.65% ~ 18.25%,0.05% ~0.08%, (11.43 ±2.10)μg/ml, P<0.05]. The G0/G1 stage proportion, CD24+ ,CD44+ ,CD24 +CD44 + cells proportion, IC50 in PANC 1 cell were ( 80. 1 ± 4.7) %, 5.31% ~ 9.84%, 72.05% ~ 93.06%,4.91% ~5.21%, (296.58±4.27) μg/ml, which were significantly higher than that those in parent cell [ (46.1 ±5.3)%, 4.09% ~4.97%, 47.71% ~55.66%, 1.48% ~2.63%, (26.17 ±3.81)μg/ml, P<0.05]. The expression of miR-590-3p in ASPC-1, PANC1 spheres was 4.67 and 4.52 times higher than the expression in parent cell lines. Conclusions Pancreatic cancer cell spheres can be isolated from ASPC-1, PANC1 by culture with SFM. miR-590-3p is up-regulated and may play an important role in regulating biological characteristics of pancreatic cancer stem cells.