CAJ | 학술논문

目的观察促血管生成素2(Ang2)单链抗体(ScFv-Ang2)对人肝癌血管生成及肿瘤生长的作用.方法体外培养人脐静脉内皮细胞(HUVEC),各组分别添加血管内皮生长因子(VEGF)、VEGF+ Ang2、VEGF+ Ang2+ ScFv-Ang2,通过HUVEC增殖测定、迁移实验、小管形成实验观察ScFv-Ang2在体外对HUVEC生物学行为的影响；建立荷人肝癌细胞株MHCC97的肝原位移植瘤模型裸小鼠24只,随机分为实验组和对照组,实验组瘤内注射ScFv-Ang2 (4×1011 pfu/ml),对照组予以生理盐水,测量肿瘤大小、肺转移灶数目、癌组织CD31蛋白表达及微血管密度(MVD)计数,观察ScFv-Ang2对肝细胞癌(HCC)血管生成及肿瘤生长的作用.结果体外实验中观察到ScFv-Ang2可抑制VEGF和Ang2诱导的HUVEC增殖、迁移及小管形成；ScFv-Ang2瘤内注射组肿瘤体积、重量、肺转移灶数目、CD31表达水平以及MVD计数均显著低于对照组(P＜0.05).结论 ScFv-Ang2在体外实验中具有抑制HUVEC形成血管的作用,体内实验中对裸鼠人肝癌具有抑制血管生成和肿瘤生长的作用.
목적 관찰촉혈관생성소2(Ang2)단련항체(ScFv-Ang2)대인간암혈관생성급종류생장적작용.방법 체외배양인제정맥내피세포(HUVEC),각조분별첨가혈관내피생장인자(VEGF)、VEGF+ Ang2、VEGF+ Ang2+ ScFv-Ang2,통과HUVEC증식측정、천이실험、소관형성실험관찰ScFv-Ang2재체외대HUVEC생물학행위적영향；건립하인간암세포주MHCC97적간원위이식류모형라소서24지,수궤분위실험조화대조조,실험조류내주사ScFv-Ang2 (4×1011 pfu/ml),대조조여이생리염수,측량종류대소、폐전이조수목、암조직CD31단백표체급미혈관밀도(MVD)계수,관찰ScFv-Ang2대간세포암(HCC)혈관생성급종류생장적작용.결과 체외실험중관찰도ScFv-Ang2가억제VEGF화Ang2유도적HUVEC증식、천이급소관형성；ScFv-Ang2류내주사조종류체적、중량、폐전이조수목、CD31표체수평이급MVD계수균현저저우대조조(P＜0.05).결론 ScFv-Ang2재체외실험중구유억제HUVEC형성혈관적작용,체내실험중대라서인간암구유억제혈관생성화종류생장적작용.
Objective To observe the effects of single-chain antibody against human angiopoietin 2 (Ang2) on the angiogenesis and growth of hepatocellular carcinoma in nude mice.Methods Human umbilical vein endothelial cells (HUVECs) were cultured and grouped in vitro.Vascular endothelial growth factor (VEGF),VEGF and Ang2,or VEGF + Ang2 + single-chain antibody (ScFv)-Ang2 was added into each group.Methyl thiazol tetrazolium (MTT) assay,Transwell assay and Matrigel assay were used to detect the effects of ScFv-Ang2 on angiogenic ability in vitro.A total of 24 nude mice bearing human hepatocellular carcinoma of MHCC97 cell line with a high metastatic potential were established and randomly divided into 2 groups.Experimental group accepted intratumoral injection of ScFv-Ang2,and saline injection group was set as placebo-control.Mice were sacrificed on the 21st day,tumor size and weight and metastases in lungs were measured,the expression of CD31 in tumor tissues was detected by using immunohistochemistry,and microvessel density (MVD) was counted.Results In vitro experiment concerning HUVECs,ScFv-Ang2 inhibited VEGF-and Ang2-induced proliferation,migration and tube formation of HUVECs.Statistical indexes including tumor weight and volume,metastases to lungs,CD31 expression and MVD counting in ScFv-Ang2 injection group were all significantly lower than in placebo-control group (P ＜ 0.05).Conclusion In addition to the inhibition of blood vessel formation of HUVECs in vitro,ScFv-Ang2 exerts significant inhibitory effect on tumor growth,angiogenesis and metastasis of human hepatocellular carcinoma in nude mice.