LY294002 decreases the proliferation of cancer stem cell-enriched spheroid cells from human hepatocel-lular carcinoma via inhibiting of PI3K-Akt signaling pathway
10.3760/cma.j.issn.1673-422X.2015.04.001
- VernacularTitle:LY294002抑制 PI3K-Akt 通路对富集肝癌干细胞细胞球增殖的影响
- Author:
Zhengwei LENG
;
Gang YANG
;
Yong LI
;
Gang SHI
- Publication Type:Journal Article
- Keywords:
Liver neoplasms;
Neoplastic stem cells;
Signal transduction;
Spheroid cells
- From:
Journal of International Oncology
2015;(4):241-244
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the impact of LY294002 on the proliferation of cancer stem cell-enriched spheroid cells from human hepatocellular carcinoma via regulating phosphatidylinositol-3-kinaseprotein kinase B(PI3K-Akt)signaling pathway. Methods The cancer stem cell-enriched spheroid cells were genera-ted by culturing HepG2 cells in serum-free medium. LY294002(10,20,30 μmol/ L),an inhibitor of PI3K-Akt signaling pathway,was used in the experimental groups,without used in the control group. The impact of LY294002 on the spheroid cells proliferation was confirmed by cell counting kit(CCK-8 kit). The expression of Akt was tested by Western blotting. The expression of PI3K-Akt signaling pathway downstream genes such as decoy receptor 3(DcR3),mammalian target of rapamycin(mTOR),B-cell lymphoma(Bcl)-2 and Cyclin D1 were tested by real-time PCR. Results 30 μmol/ L LY294002 could inhibit the proliferation of spheroid cells, and significant difference in the absorbance(A value)was observed between the experimental group and control group[(0. 14 ± 0. 03)vs(0. 56 ± 0. 01),t = - 8. 915,P = 0. 000]. The expression level of phosphorylated Akt protein increased[(0. 57 ± 0. 08)vs(0. 16 ± 0. 42),t = 6. 027,P = 0. 026]. The mRNA of DcR3 [(0. 38 ± 0. 08)vs 1,t = 13. 060,P = 0. 006],mTOR[(0. 37 ± 0. 04)vs 1,t = 30. 363,P = 0. 001],Bcl-2 [(0. 26 ± 0. 04)vs 1,t = 33. 554,P = 0. 001]and Cyclin D1[(0. 10 ± 0. 02)vs 1,t = 63. 528,P = 0. 000] decreased. Conclusion LY294002 could inhibit the proliferation of cancer stem cell-enriched spheroid cells from human hepatocellular carcinoma via inhibiting PI3K-Akt signaling pathway.
