期刊文献+

EphA2调控头颈部鳞状细胞癌血管新生及转移的体内实验 被引量:3

EphA2 promotes angiogenesis and metastasis of head and neck squamous cell carcinoma in vivo
原文传递
导出
摘要 目的 研究EphA2对头颈部鳞状细胞癌(简称鳞癌)血管新生及颈淋巴转移的影响.方法 利用慢病毒介导的短发夹RNA( short hairpin RNA,shRNA)沉默EphA2在高转移性头颈鳞癌细胞株M2中的表达,嘌呤霉素筛选稳定克隆,反转录PCR及Western blot技术检测EphA2沉默效果;构建头颈鳞癌高转移动物模型,通过HE染色验证其成瘤及颈淋巴转移情况;免疫组织化学技术检测移植瘤微血管密度,Western blot技术检测移植瘤标本中EphA2及血管内皮生长因子(VEGF)的表达情况.结果 筛选出EphA2基因稳定沉默的M2细胞(定义为M2 EphA2 RNA+即实验组),并成功构建头颈鳞癌高转移动物模型,成瘤率达100%.25 d后,实验组移植瘤体积为(430.7±190.0) mm3(x±s,以下同)较对照组的(1179.0 ±289.4) mm3明显减少(t =5.597,P<0.01),质量显著减轻(t=-4.560,P<0.05),且双侧颈淋巴转移率明显降低(Mann-Whitney U=10.0,P<0.05).Western blot 显示实验组移植瘤组织中EphA2及VEGF蛋白表达显著下调,免疫组织化学技术发现实验组瘤体微血管密度为(4.74±0.67)个/视野,显著少于对照组的(14.17±0.59)个/视野(t=26.751,P<0.01).结论 EphA2沉默能抑制头颈部鳞癌细胞的生长及转移,并能明显减少头颈部鳞癌组织内的肿瘤新生血管,且该肿瘤新生血管的抑制可能与促血管生成因子VEGF的减少有关. 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) mm3 ((x) ± s) vs(1179.0±289.4) mm3] and weight [(0.26-±0.10) gvs (0.54±0.12) g] (bothP〈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.
出处 《中华耳鼻咽喉头颈外科杂志》 CAS CSCD 北大核心 2012年第1期53-57,共5页 Chinese Journal of Otorhinolaryngology Head and Neck Surgery
基金 国家自然科学基金(81172558,81071757,30901664,30872852) 高等学校博士学科点专项科研基金课题(20100162110036,20090162110065) 湖南省科技厅重点项目(2010TP4012-1) 湖南省研究生学位论文创新基金(CX201113059)
关键词 头颈部肿瘤 鳞状细胞 受体 EPHA2 肿瘤转移 新生血管化 病理性 Head and neck neoplasms Carcinoma, squamous cell Receptor, EphA2 Neoplasms metastasis Neovascularization, pathologic
  • 相关文献

参考文献4

二级参考文献41

  • 1王彦君,孔维佳,熊新高,张松,孙大为.Survivin和VEGF蛋白在喉癌组织中的表达及其相关性研究[J].临床耳鼻咽喉科杂志,2005,19(18):838-841. 被引量:10
  • 2上官翰京,李志春,张晖萍,周爱东.喉鳞状细胞癌中RAGE与amphoterin的表达[J].山东大学耳鼻喉眼学报,2006,20(3):204-205. 被引量:3
  • 3Chin D,Boyle GM,Porceddu S,et al.Head and neck cancer:past,present and future.Expert Rev Anticancer Ther,2006,6:1111-1118.
  • 4Zender CA,Petruzzelli GJ.Why do patients with head and neck squamous cell carcinoma experience distant metastases:can they be prevented? Curr Opin Otolaryngol Head Neck Surg,2005,13:101-104.
  • 5Almadori G,Bussu F,Cadoni G,et al.Muhistep laryngeal carcinogenesis helps our understanding of the field cancerisation phenomenon:a review.Eur J Cancer,2004,40:2383-2388.
  • 6Rogers SJ,Harrington KJ,Rhys-Evans P,et al.Biological significance of c-erbB family oncogenes in head and neck cancer.Cancer Metastasis Rev,2005,24:47-69.
  • 7Detre S,Saclani Jotti G,Dowsett M.A "quickscore" method for immunohistochemical semiquantitation:validation for cestrogen receptor in breast carcinomas.J Clin Pathol,1995,48:876-878.
  • 8Liang X,Yang D,Hu J,et al.Hypoxia inducible factor-alpha expression correlates with vascular endothelial growth factor-C expression and lymphoangiogenesis/angiogenesis in oral squamous cell earcinoma.Anticancer Res,2008,28:1659-1666.
  • 9Weidner N,Folkman J,Pozza F,et al.Tumor angiogenesis:a new significant and independent prognostic indicator in early-stage breast carcinoma.J Natl Cancer Inst,1992,84:1875-1887.
  • 10Zhong H,De Marzo AM,Laugtner E,et al.Overexpression of hypoxia-inducible factor-1 alpha in common human cancers and their metastases.Cancer Res,1999,59:5830-5835.

共引文献37

同被引文献15

  • 1Fayette J, Montella A, Chabaud S, et al. Paclitaxel is effec- tive in relapsed head and neck squamous cell carcinoma: a ret- rospective study of 66 patients at a single institution [ J ]. Anti- cancer Drugs, 2010, 21 (5): 553-558.
  • 2Holleman A, Chung I, Olsen RR, et al. miR-135a contrib- utes to paclitaxel resistance in tumor cells both in vitro and in vivo [J]. Oncogene, 2011, 30(43): 4386 -4398.
  • 3Ireton RC, Chen J. EphA2 receptor tyrosine kinase as a prom- ising target for cancer therapeutics [ J ]. Curt Cancer Drug Tar- gets, 2005, 5(3): 149 -157.
  • 4Landen CN, Merritt WM, Mangala LS, et al. Intraperitoneal delivery of liposomal siRNA for therapy of advanced ovarian cancer[J]. Cancer Biol Ther, 2006, 5 (12) : 1708 - 1713.
  • 5Landen CN, Jr, Lu C, Han LY, et al. Efficacy and antivas- cular effects of EphA2 reduction with an agonistic antibody in o- varian cancer[J]. J Natl Cancer Inst, 2006, 98 (21): 1558 - 1570.
  • 6Kavallaris M. Mierotubules and resistance to tubulin-binding a- gents[J]. Nat Rev Cancer, 2010, 10(3) : 194 -204.
  • 7Liu Y, Zhang X, Qiu Y, et al. Clinical significance of E- phA2 expression in squamous-eell carcinoma of the head and neck[J]. J Cancer Res Clin Oneol, 2011, 137(5): 761 -769.
  • 8Liu Y, Yu C, Qiu Y, et al. Downregulation of EphA2 ex- pression suppresses the growth and metastasis in squamous-cell carcinoma of the head and neck in vitro and in vivo [ J ] . J Cancer Res Clin Oncol, 2012, 138(2): 195-202.
  • 9Tan P, Liu Y, Yu C, et al. EphA2 silencing in nasopharyn- gem carcinoma leads to decreased proliferation, invasion and sensitization to Paclitaxel [ J]. Oncol lett, 2012, 4 (3) : 429 -434.
  • 10Rosenfeldt MT, Ryan KM. The multiple roles of autophagy in cancer [ J ]. Carcinogenesis, 2 011 , 3 2 ( 7 ) : 9 5 5 - 9 6 3.

引证文献3

二级引证文献10

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部