title: [EphA2 promotes angiogenesis and metastasis of head and neck squamous cell carcinoma in vivo].
abstract: OBJECTIVE
  To investigate the effects of EphA2 on the angiogenesis and cervical lymph node metastasis of squamous cell carcinoma of the head and neck (SCCHN) in vivo.
  
  
  METHODS
  EphA2 short hairpin (shRNA) lentiviral particles were used to knockdown the expression of EphA2 in SCCHN cell line M2 with high lymph nodes metastasis rate. Stable clones, obtained by puromycin screening, were assayed by RT-PCR and Western blot to validate the gene silencing efficiency and were used to establish SCCHN metastatic xenograft mouse model. Hematoxylin-eosin staining was applied to identify cervical lymph node metastasis of SCCHN in xenografted tumors. Immunohistochemistry was used to observe microvessel density. Western blot was used to investigate the protein expressions of EphA2 and vascular endothelial, growth factor (VEGF).
  
  
  RESULTS
  EphA2 shRNA lentiviral particles efficiently decreased the mRNA and protein expressions of EphA2 in SCCHN cell line M2, which were further successfully utilized to establish SCCHN metastatic xenograft mouse model. Compared with xenografted tumors in control group, xenografted tumors in M2EphA2RNAi(+) group decreased significantly tumor volume [(430.7 ± 190.0) mm(3) (x(-) ± s) vs (1179.0 ± 289.4) mm(3)] and weight [(0.26 ± 0.10) g vs (0.54 ± 0.12) g] (both P < 0.05). More importantly, bilateral cervical lymph node metastasis rate in M2EphA2RNAi(+) was also greatly declined (Mann-Whitney U = 10.0, P < 0.05). Decreased protein expressions of EphA2 and VEGF and microvessel density were observed in M2EphA2RNAi(+) group (t = 26.751, P < 0.01).
  
  
  CONCLUSIONS
  Knockdown of EphA2 expression led to the inhibition of tumor growth and metastasis in SCCHN nude mouse model. More importantly, SCCHN angiogenesis was also impeded, which might be associated with the decreased expression of VEGF.
authors.affiliation: Department of Otorhinolaryngology Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.forename: Chang-yun
authors.initials: CY
authors.lastname: Yu
authors.forename: Yuan-zheng
authors.initials: YZ
authors.lastname: Qiu
authors.forename: Dong-hai
authors.initials: DH
authors.lastname: Huang
authors.forename: Xiao-juan
authors.initials: XJ
authors.lastname: Zhou
authors.forename: Ping-qing
authors.initials: PQ
authors.lastname: Tan
authors.forename: Jian-yun
authors.initials: JY
authors.lastname: Xiao
authors.forename: Yong-quan
authors.initials: YQ
authors.lastname: Tian
mesh_terms: D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D045744:Cell Line, Tumor; D020868:Gene Silencing; D006258:Head and Neck Neoplasms / Q000473:pathology*; D006801:Humans; D008207:Lymphatic Metastasis; D051379:Mice; D008819:Mice, Nude; D009389:Neovascularization, Pathologic; D011379:Prognosis; D034741:RNA, Small Interfering; D036104:Receptor, EphA2 / Q000235:genetics*; D000077195:Squamous Cell Carcinoma of Head and Neck
chemical_list: D034741:RNA, Small Interfering; D036104:Receptor, EphA2