摘要
目的探讨自行研制的肝素酶多肽抗体对人肝癌HCCLM6细胞生长和侵袭的影响。方法采用二苯基溴化四氮唑蓝（MTT）法检测细胞生长曲线，平板克隆形成试验检测细胞集落数，流式细胞仪检测细胞周期，通过肝素酶活性测定和Transwell侵袭小室试验，了解自行研制的3种肝素酶多肽兔多克隆抗体（P1、P2和P3多肽抗体）对HCCLM6肝癌细胞牛长及侵袭的影响。结果与正常兔IgG对照组比较，P1、P2和P3多肽抗体对HCCLM6肝癌细胞的生长、细胞周期及增殖活性均无明显影响。终浓度为100μg／ml的P1、P2多肽抗体与HCCLM6细胞共培养后，肝素酶活性分别降低42.9％和39.1％（P〈0.01），Transwell侵袭小室中48h穿膜细胞数明显减少，抑制率分别为52.5％和36.6％（P〈0.05）。结论肝素酶P1和P2多肽抗体能抑制人肝癌HCCLM6细胞的肝素酶活性和侵袭能力，P3多肽抗体对肝癌细胞的肝素酶活性和侵袭能力无影响；肝素酶P1、P2和P3多肽抗体对HCCLM6细胞的生长、细胞周期和增殖能力均无明显影响。
Objective To investigate the effects of the self-developed anti-heparanase polypeptide antibodies on growth and invasion of human hepatocellular carcinoma HCCLM6 cells. Methods Using MTT, flow cytometry, plate clone formation, transwell invasion and heparan degrading enzyme assay, the growth and invasion changes of human hepatocellular carcinoma HCCLM6 cells by co-culture with each of three self-developed rabbit anti-heparanase polyclonal antibodies were detected. Results Compared with normal rabbit IgG, in the presence of each anti-heparanase polypeptide antibody, the growth, cell cycle and clone formation remained unchanged, and under the P1 or P2 anti-heparanase polypeptide antibody （with final concentration 100μg/ml） , the cell invasiveness was inhibited by 52.5% and 36.6%, respectively, and the heparanase activity was inhibited by 42.9% and 39.1%, respectively. Conclusion The P1 and P2 antiheparanase polypeptide antibodies can effectively inhibit the invasion ability and heparanase activity of liver cancer HCCLM6 cells. However, All the three antibodies have no effects on its growth, cell cycle and clone formation.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2009年第1期10-14,共5页
Chinese Journal of Oncology
基金
国家自然科学基金（30570816）.
关键词
肝素酶
多肽抗体
肝肿瘤
增殖
侵袭
Heparanase
Polypeptide antibody
Hepatocellular carcinoma
Proliferation
Invasion
