Impacts of hypoxia on the features and chemoresistance of cancer stem cells in Hep-2 cells and underlying mechanism
10.3760/cma.j.issn.1673-0860.2012.03.011
- VernacularTitle:低氧对Hep-2细胞肿瘤干细胞特性和化疗抵抗的影响及其作用机制
- Author:
Yong-Tao QU
1
;
Xiao-Ming LI
;
Ou XU
;
Mao-Xin WANG
;
Xiu-Ying LU
Author Information
1. 河北医科大学
- Keywords:
Laryngeal neoplasms,Carcinoma,squamous cells;
Anoxia;
Neoplastic stem cells;
Hypoxia-inducible factor 1,alpha subunit
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2012;47(3):228-233
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of hypoxia on the features and chemoresistance of cancer stem cells in Hep-2 cells and underlying mechanism. Methods The shRNA interference recombinant plasmid targeting HIF-1α was synthesized and transfected into Hep-2 cells. The HIF-1αknockdown Hep-2 cells were established after clonal selection and the expression of HIF-1α was measured.The cellular features including proliferation,clonal formation,cell cycle,apoptosis and CD133 phenotype were measured in Hep-2 cells cultured under hypoxic condition in vitro. CD133 + cells were sorted from Hep-2 cells with flow cytometry. Clonal formation test and cisplatin treatment were carried out,and theexpressions of related genes( Oct-4,suvivin and p53 ) in CD133 + cells were measured.Results HIF-1αknockdown Hep-2 cells was successfully established,as evidenced by the reduced mRNA and proteinexpressions of HIF-lα. The Hep-2 cells cultured under hypoxic microenvironment showed higher proliferation and clonal formation activity,cell cycle arrest in G0/G1,lower apoptosis,up-regulated CD133,however the effects of hypoxia reduced in HIF-1α knockdown Hep-2 cells.CD133 + cells were successfullysorted from Hep-2 cells,and the CD133+ cells showed increased clonal formation activity and cisplatintreatment resistance in hypoxia. Also the effects of hypoxia on CD133 + cells decreased with HIF-1αknockdown,showing down-regulated Oct-4 and survivin and up-regulated p53.Conclusions Hypoixa caninduce the features of cancer stem cells in Hep-2 cells and increase proliferation,differentiation and chemoresistant ability of CD133 + cells,which might be correlated with the changes in expressions of HIF-1 αand related genes regulated by HIF-1 α.
