蚯蚓纤溶酶抗肝癌转移作用的实验研究  

Anti-metastasis Activity of Earthworm Fibrinolytic Enzyme on Hepatoma Cell in Vivo

作　　者：常春霞[1] 陈洪[1] 季红[2] 张治国[3] 

机构地区：[1]东南大学附属中大医院消化科,南京210009 [2]东南大学临床医学院,南京210009 [3]南京农业大学生命科学院免疫生化研究所,南京210095

出　　处：《中药新药与临床药理》2009年第6期520-524,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：江苏省中医药局科技基金(HL07067);东南大学科技基金(KJ0790295)

摘　　要：目的探讨蚯蚓纤溶酶(earthworm fibrinolytic enzyme,EFE)对人肝癌细胞转移的影响。方法建立裸鼠人肝癌高转移原位移植瘤模型,造模后7d将裸鼠随机分为模型对照组和EFE高、低剂量组,共3组,每组7只,每天分别给予生理盐水和1600,800uku/kgEFE灌胃,连续30d。实验结束时完整剥除种植瘤并称重;肉眼直接观察种植瘤的肝内播散及腹腔种植情况,计算肝内播散率及腹腔种植率;通过病理切片观察肺转移肿瘤灶的数目;采用RT-PCR及Western blot方法检测移植瘤中FAK(focus adhesion kinase)及β1-整合素蛋白的表达。结果与模型对照组相比,EFE低、高剂量组原位种植瘤瘤重减轻(P<0.05或P<0.01);种植瘤肝内播散率和腹腔种植率降低,其中,EFE高剂量组与模型对照组比较,具有显著性差异(P<0.05);肺转移灶数目明显减少,与模型对照组相比,具有显著性差异(P<0.05或P<0.01)。在原位种植瘤和移植瘤中FAK及β1-整合素在mRNA及蛋白水平的表达方面,EFE高、低剂量组的表达均明显下降,与模型对照组比较具有非常显著性差异(P<0.01)。结论EFE具有抗肝癌转移的作用,其机理可能与EFE能够抑制FAK及β1-整合素的表达有关。Objective To investigate the inhibitory effect of earthworm fibrinolytic enzyme （EFE） on the metastasis of human hepatoma cells. Methods A metastatic model of human hepatocellular carcinoma in nude mice via orthotopic iraplantation was established. After the modeling for 7 days, the dose EFE group （800 uku/kg · d^- 1）, and high-dose EFE days, the implanted tumors were we mice were divided randomly into the control group, low - group （1600 uku/kg · d^-1）. After administration for 30 ted, and the intrahepatic transmission rate and abdominal cavity seeding late were calculated after examination by naked eyes. Then the puhnonary metastasis were examined under microscope, and the expression of focus adhesion kinase （FAK） and [31 - intigrin were detected through RT - PCR and western blotting method. Results Compared with the model control group, the mean weight of the orthotopic tumor was reduced, and the intrahepatic transmission rate and abdominal cavity seeding rate were decreased in FEF groups （ P 〈 0. 05 or P 〈 0.01 ）, the difference of the above indexes being significant between high - dose EFE group and the control group （ P 〈 0.05）. The number of pulmonary metastasis focuses was reduced （ P 〈 0.05 or P 〈 0.01 ）, and mRNA and protein expression levels of FAK and β1 -intigrin were decreased （P 〈 0.01） as compared with those in the model control group. Conclusion EFE has an effect on counteracting the metastasis of human hepatocellular carcinoma, which may be related to the depression of FAK and β1 - integrin expression.

关 键 词：蚯蚓纤溶酶 肝癌 转移 原位移植瘤模型 裸鼠 

分 类 号：R285.5[医药卫生—中药学]