title: [Inhibitory effect of full-length spleen tyrosine kinase on invasion and metastasis of human laryngeal squamous cells].
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). Transwell 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) mm(3) tumor formed from transfected-Sky (L) cells in nude mice, was less than those from Hep-2-neo cells (750.77 ± 40.83) mm(3) and Hep-2 cells (770.77 ± 30.83) mm(3), with statistically significant difference (F = 165.78, P < 0.01).
  
  
  CONCLUSIONS
  Down-regulation 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.
authors.affiliation: Department of Otorhinolaryngology, Taizhou Municipal Hospital, Taizhou 318000, China. Email: li00hai@sohu.com.
authors.forename: Zhihai
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Otorhinolaryngology, Taizhou Municipal Hospital, Taizhou 318000, China.
authors.forename: Zhiyi
authors.initials: Z
authors.lastname: Cai
authors.affiliation: Department of Otorhinolaryngology, Taizhou Municipal Hospital, Taizhou 318000, China.
authors.forename: Baohong
authors.initials: B
authors.lastname: Tao
authors.affiliation: Department of Otorhinolaryngology, Taizhou Municipal Hospital, Taizhou 318000, China.
authors.forename: Qiaozhi
authors.initials: Q
authors.lastname: Jin
mesh_terms: D000818:Animals; D002294:Carcinoma, Squamous Cell; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015536:Down-Regulation; D004847:Epithelial Cells; D015316:Genetic Therapy; D005822:Genetic Vectors; D006258:Head and Neck Neoplasms; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D007822:Laryngeal Neoplasms / Q000378:metabolism*; D007830:Larynx; D007831:Larynx, Artificial; D051379:Mice; D008819:Mice, Nude; D016609:Neoplasms, Second Primary; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000077195:Squamous Cell Carcinoma of Head and Neck; D000072377:Syk Kinase; D014162:Transfection
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C000606897:Syk protein, mouse