突变型K5重组蛋白抑制小鼠肝癌血管生成和肿瘤生长的作用  

Plasminogen kringle 5 deletion mutantⅠ inhibits the neovascularization and growth of the mouse hepatocarcinoma

作　　者：杨霞[1] 王青松[1] 程锐[1] 李朝阳[2] 蔡卫斌[1] 杨中汉[1] 李民友 何光耀[1] 高国全[1] 

机构地区：[1]中山大学基础医学院生物化学教研室,广东广州510080 [2]中山大学中山眼科中心,广东广州510080 [3]广州市启源生物科技有限公司,广东广州510630

出　　处：《中国病理生理杂志》2008年第2期220-224,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30370313No.30570372No.30600724);中华医学基金会资助项目(CMB-SUMS学者项目98-677);教育部新世纪人才计划资助项目(No.NCET-04-0792);广东省自然科学基金研究团队资助项目(No.06201946);广东省科技计划项目重大专项资助项目(No.2005A10902003);广东省医学科研基金资助项目(No.A2006175);广州市科技攻关重点基金资助项目(No.2005Z3-E4031No.2006Z3-E4111)

摘　　要：目的:体内实验观察人纤溶酶原K5缺失突变体Ⅰ(K5 mut1)对HepA小鼠肝癌血管生成和肿瘤生长的影响。方法:大肠杆菌中表达,组氨酸结合柱亲和层析、纯化获得K5 mut1蛋白,SDS-PAGE和Western blot-ting方法鉴定其表达;建立皮下种植肝癌小鼠模型,腹腔注射不同剂量K5 mut1重组蛋白,检测抑瘤率及肝癌组织微血管密度(MVD)。结果:SDS-PAGE及Western blotting鉴定获得K5 mut1纯化蛋白;K5 mut1剂量依赖性地抑制小鼠肝癌(HepA)实体瘤生长,不同剂量K5 mut1治疗组肝癌组织MVD低于对照组,并随K5 mut1用药剂量增加而降低。结论:K5 mut1具有抑制小鼠肝癌生长的作用,抑制肿瘤血管生成可能是K5 mut1抑制肿瘤生长的主要机制。结果提示K5 mut1具有治疗肝癌的潜在临床价值。AIM ： To investigate the effects of plasminogen kringle 5 （ K5 ） deletion mutant Ⅰ （ K5 mut 1 ） on the neovascularization and growth of hepatocarcinoma in vivo. METHODS： K5 mut1 was expressed in E. colt BL -21 （ DE3 ） induced by IPTG and purified by Ni^2＋ - His Bind Resin affinity chromatography. The purity and identity of K5 mut1 was examined by SDS - PAGE and Western blotting analysis. K5 mut1 activity was evaluated in vivo, HepA - grafed hepatocarcinoma mouse model was established by subcutaneously injection of mouse hepatoma cells（ 1 × 10^6） into the oxter of mice, and the mice were then divided into different groups and treated with PBS and K5 mut1 at different doses, respectively. The tumor suppressing rate and micro vascular density （MVD） in hepatoma tissues were assayed. RESULTS： The purity of recombinant K5 mut1 was over 90% according to the analysis of SDS - PAGE and Western blotting analysis. K5 mut1 significantly inhibited the growth of tumor in a HepA - grafed hepatocarcinoma mouse model and decreased microvessel density （MVD） in hepatoma tissues in a dose -dependent manner, compared with PBS control group. CONCLUSION： K5 mut1 exhibits anti - tumor effect in a HepA - grafted hepatocarcinoma mice model by the anti - angiogenic activity. These results support the conclusion that K5 mut1 may be a promising angiogenesis inhibitor and tumor suppressor with considerable therapeutic potential in liver cancer.

关 键 词：纤维蛋白溶酶原 突变 癌 肝细胞 新生血管化 病理性 

分 类 号：Q784[生物学—分子生物学]