Inhibitory effect of full-length spleen tyrosine kinase on invasion and metastasis of human laryngeal squamous cells
10.3760/cma.j.issn.1673-0860.2014.11.014
- VernacularTitle:全长型脾酪氨酸激酶基因转染对喉癌细胞恶性生物学行为的抑制作用
- Author:
Zhihai LI
1
;
Zhiyi CAI
;
Baohong TAO
;
Qiaozhi JIN
Author Information
1. 318000,浙江省台州市立医院耳鼻咽喉科
- Keywords:
Laryngeal neoplasms;
Protein-tyrosine kinases;
Transfection;
Cell line,tumor
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2014;49(11):943-949
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effect of gene transfection of full length spleen tyrosine kinase (Syk (L)) on the biological behavior of malignant cancer cells.Methods Eukaryotic expression vector pIRES2-EGFP-Syk (L) was constrauted and sequenc.Laryngeal carcinoma cell line Hep-2 were transfected with pIRES2-EGFP-Syk (L) vectors or blank vectors.The expressions of mRNA and protein were examined by real time fluorescence quantitative polymerase chain reaction (Q-RT-PCR) and Western blot analysis.CCK-8 method was used for evaluating cell proliferation,Transwell for cell invasion capacity in vitro,and tumor formation in nude mice for in vivo tumorigenicity.Results pIRES2-EGFP-Syk (L) vectors were successfully construct and transfected to Hep-2 cells.Q-PCR showed that mRNA expression level in Hep-2 cells transfected with Sky (L) (28.395 ± 0.067) was higher than those in Hep-2-neo cells transfected with blank vectors (3.891 ± 0.021) and Hep-2 cells with no transfection (1.005 ± 0.012),with statistically significant difference (F =104.02,P ＜ 0.01).Western blot showed that protein expression level of transfected-Sky (L) cells (0.821 ± 0.047) was significantly higher than those of Hep-2-neo cells (0.558 ±0.031) and Hep-2 cells (0.468 ± 0.031),and the difference was statistically significant (F =112.32,P ＜ 0.01) ; CCK-8 assay showed OD value (1.390 ± 0.067) of transfected-Sky (L) cells was lower than those of Hep-2-neo cells (1.830 ± 0.067) and Hep-2 cells (1.920 ± 0.040),and the difference was statistically significant (F =107.64,P ＜ 0.01).Transwe11 assay showed average cell number per field of transfected-Sky (L) cells (176.04 ± 22.32) was higher than those of Hep-2-neo cells (301.02 ± 21.45) and Hep-2 cells (336.04 ± 26.01) with statistically significant difference (F =123.46,P ＜0.01).The volume (250.77 ± 34.83) mm3 tumor formed from transfected-Sky (L) cells in nude mice,was less than those from Hep-2-neo cells (750.77 ± 40.83) mm3 and Hep-2 cells (770.77 ±30.83) mm3,with statistically significant difference (F =165.78,P ＜0.01).Conclusion Downregulation of Syk in Hep-2 cells is associated with the malignant biological behaviors of the cells.Syk (L)may be a potential target in gene therapy for laryngeal squamous cell carcinoma.
