反式转录激活因子-氨基端24个氨基酸穿膜融合多肽对结直肠癌血管形成的影响  

Effect of cell-permeable trans-activator of transcription-N24 fusion peptide on angiogenesis of colorectal cancer

作　　者：杨熹[1] 李海杰[1] 王桂华[1] 陈成[1] 杨锐[1] 龚建平[1] 胡俊波[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤研究所/胃肠外科,430030武汉

出　　处：《中华实验外科杂志》2014年第2期239-241,共3页Chinese Journal of Experimental Surgery

基　　金：国家973计划资助项目（2009CB521802）;国家自然科学基金资助项目（30872472、30973496、30800569）;湖北省自然科学基金资助项目（2008CDB174、2009CDB239）

摘　　要：目的 观察反式转录激活因子-氨基端24个氨基酸（TAT-N24）穿膜融合多肽对结直肠癌血管形成的影响.方法 对结直肠癌细胞Lovo过表达p55PIK和/或同时给予p55PIK的特异性抑制剂TAT-N24后,通过Western blot法检测乏氧诱导因子-1α的表达,通过双荧光素酶报告系统检测血管内皮生长因子-A的启动子活性,并用酶联免疫吸附试验检测其蛋白分泌,最后通过体外血管形成实验检测p55PIK及TAT-N24对结直肠癌血管形成的影响.结果 过表达p55PIK促进结直肠癌细胞株Lovo中乏氧诱导因子-1α的表达,进而增强血管内皮生长因子-A的启动子活性（p55PIK组比对照组,常氧：9.1±1.1比1.0±0.2,乏氧：14.5 ±1.6比1.0±0.3）,并且促进其在培养基上清中的分泌[p55PIK组比对照组,常氧：（412±23） mg/L比（150±18） mg/L,乏氧：（859±203） mg/L比（233±12） mg/L],而且其条件培养基可促进脐静脉内皮细胞形成血管,而TAT-N24可拮抗上述过程,抑制结直肠癌的血管形成.结论 p55PIK可促进结直肠癌的血管形成,而TAT-N24作为其特异性抑制剂,可抑制结直肠癌的血管形成.Objective To investigate the effect of cell-permeable trans-activator of transcriptionN24 （TAT-N24） fusion peptide on angiogenesis of colorectal cancer Lovo cell line.Methods Lovo cells were transfected with plasmids for overexpression of p55PIK,with or without treatment with TAT-N24,a specific inhibitor of p55PIK,for the indicated time,then the expression of hypoxia inducible factor-1 was detected by using Western bloting,and the transcriptional activity of promoter of vascular growth factor-α was measured through Dual-Luciferase Reporter Assay.Meanwhile,enzyme linked immunosorbent assay （ELISA） was performed to detect the expression of vascular growth factor-α,and in vitro angiogenesis assay was conducted by tube formation.Results The expression of hypoxia inducible factor-1 and vascular growth factor-α was both elevated [p55PIK vs control,for normoxia （412 ±23） mg/L vs.（150-± 18） mg/L,and for hypoxia （859 ±203） mg/L vs.（233 ± 12） mg/L],and the transcriptional activity of promoter of vascular growth factor-α （p55PIK vs control,for normoxia 9.1 ± 1.1 vs.1.0 ± 0.2,and for hypoxia 14.5 ± 1.6vs.1.0 ±0.3） and the tube formation of human umbilical vein endothelial cell （HUVEC） was both enhanced under the overexpression of p55PIK.Meanwhile,TAT-N24 could antagonize these effects of p55PIK on hypoxia inducible factor-1 and vascular growth factor-a,and inhibit the angiogenesis of colorectal cancer.Conclusion p55PIK could enhance the angiogenesis of colorectal cancer,and meanwhile,TAT-N24,as a specific inhibitor of p55PIK,effectively antagonize the functions of p55PIK and inhibit the angiogenesis of colorectal cancer.

关 键 词：结直肠癌 p55PIK 反式转录激活因子 血管形成 血管内皮生长因子 

分 类 号：R735.3[医药卫生—肿瘤]