Establishment of CoCl2 induced hypoxic cell model of oral squamous cell carcinoma and a preliminary study on the biological behaviour change
10.3760/cma.j.issn.1002-0098.2015.03.012
- VernacularTitle:氯化钴诱导建立口腔鳞状细胞癌低氧模型及其生物学行为初探
- Author:
Shengwei HAN
1
;
Wei HAN
;
Yanhong NI
;
Zhiyong WANG
;
Qingang HU
Author Information
1. 210008,南京大学医学院附属口腔医院·南京大学口腔医学院口腔颌面外科
- Keywords:
Oral sprays;
Hypoxic cell model;
CoCl2
- From:
Chinese Journal of Stomatology
2015;50(3):173-177
- CountryChina
- Language:Chinese
-
Abstract:
Objective To mimic oral squamous cell carcinoma(OSCC) cell hypoxia by using chemical agent CoCl2 and to investigate its biological behaviour.Methods Oral squamous cell carcinoma cell lines HSC-3 and SCC-4 were exposed to different concentration of CoCl2.HSC-3 and SCC-4 cells were treated with 50,100,150,200 μ mol/L CoCl2.Expression of hypoxia inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF) and B-cell lymphoma-2(BCL-2) were measured by real time polymerase chain reaction(PCR) and Western blotting in both mRNA and protein level.Cell proliferation,cell apoptosis and cell cycle were detected to analyze its biological behaviour.Both wound healing and Transwell assay were applied to test the ability of cell igration.Results The result showed that after treatment of 150 μ mol/L CoCl22 for 24 h,mRNA level of HIF-1α,VEGF and Bcl-2 was increased by 6.00±0.20,5.40±0.40,5.40±0.30 (SCC-4); 5.60±0.30,5.20±0.60,5.80±0.40(HSC-3).OSCC cells treated with 150 μmol/L CoCl2 for 24 h were collected.Compared with control group,the growth rate of cells was significantly decreased,P value was less than 0.05 (when HSC-3,SCC-4 cultured for 2 and 3 days).The apoptosis of OSCC cells was increased when treated with 150 μmol/L CoCl2 for 24 h:HSC-3 2.25％(control group) and 5.82％(treatment group); SCC-4 2.58％(control group) and 10.27％ (treatment group).The migration ablility of OSCC cells was decreased when using 150 μmol/L CoCl2 for 24 h.The migration area ratio was(31.5±2.3) ％(HSC-3),(29.1± 1.5) ％ (SCC-4) in control group and(18.3± 1.9) ％ (HSC-3),(13.2±0.8)％ (SCC-4) in treatment group(P＜0.05).Conclusions The hypoxic cell model of OSCC could be induced by CoCl2.The expression level of hypoxic markers was up regulated significantly and the cells biological behaviour changed including decreased cell proliferation,increased apoptosis and decreased migration.
