重组去整合素rAdinbitor对肝癌细胞H22生长的抑制作用

Inhibitory Effect of Recombinant Disintegrin rAdinbitor on the Growth of H22 Hepatoma Cells

目的:探讨源于白眉蝮蛇的重组去整合素rAdinbitor对肝癌细胞H22生长的抑制作用及其机制。方法:通过诱导重组表达、纯化、鉴定等步骤获得rAdinbitor;采用右腋皮下注射1×106个H22细胞制备肝癌小鼠模型,随机分为模型组(0.9%氯化钠注射液0.2 ml/kg)、阳性对照组(重组人血管内皮抑制素1.65 mg/kg)和高、中、低剂量治疗组(rAdinbitor 5、1.25、0.5 mg/kg),每组8只,肌肉注射相应药物,每日1次,连续给药3周后处死小鼠,取瘤称质量,镜下观察肿瘤组织切片中5个视野下的血管数量。结果:成功诱导表达并纯化rAdinbitor,其蛋白含量为1.4 mg/ml,相对分子质量约为9 kD。各组小鼠瘤质量和血管数量分别为模型组:(1.513±0.922)g和3.34±0.58;阳性对照组:(0.562±0.304)g和1.33±0.58;高剂量治疗组:(0.318±0.205)g和1.18±0.56;中剂量治疗组:(0.434±0.323)g和1.32±0.60;低剂量治疗组:(0.536±0.328)g和1.35±0.58。与模型组比较,各剂量治疗组小鼠的瘤质量和血管数量均明显下降(P<0.05);与阳性对照组比较,中、高剂量治疗组小鼠的瘤质量差异有统计学意义(P<0.05),高剂量治疗组小鼠的血管数量差异有统计学意义(P<0.05),其余差异无统计学意义。结论:rAdinbitor对H22细胞的生长具有明显的抑制作用,其机制可能是抑制瘤组织周边新血管的形成。

OBJECTIVE： To investigate the inhibitory effect and mechanism of recombinant disintegrin rAdinbitor from Gloydi- us blomhoffi brevicaudus on the growth of H22 hepatoma cells. METHODS ： rAdinbitor was obtained after recombinant expression, purification and identification. Hepatic cancer model was induced by 1 x 106 H22 cells subcutaneously via right armpit. Model mice were randomly divided into model group （0.9% Sodium chloride injection 0.2 ml/kg）, positive control group （recombinant human endostatin 1.65 mg/kg） and high-dose, medium-dose and low-dose groups （rAdinbitor 5, 1.25 and 0.5 mg/kg） with 8 rats in each group. They were given relevant drugs intramuscularly once a day for consecutive 3 weeks. Mice were sacrificed and tumors were weighted. The vescular number of 5 visual fields was counted under the microscope. RESULTS： The total protein reached 1.4 mg/ml and relative molecular mass was 9 kD after rAdinbitor was induced and purified successfully. The tumor weight and vascular number of each group were （1.513 ± 0.922） g and 3.34 _± 0.58 in model group, （0.562 ± 0.304） g and 1.33 ± 0.58 in positive control group, （0.318 ± 0.205） g and 1.18 ± 0.56 in high-dose group, （0.434 ± 0.323） g and 1.32 ± 0.60 in medium-dose group and （0.536 ± 0.328） g and 1.35± 0.58 in low-dose group. Compared with model group, tumor weight and vascular number of other groups were decreased significantly （P〈0.05） ; compared with positive control group, there was statistical significance in tumor weight of medium-dose and high-dose groups （P〈0.05）. There was statistical significance in vascular number of high-dose group （P〈0.05）; there was no other statistical significance. CONCLUSIONS： The growth of H22 hepatoma cells is inhibited by rAdinbi- tor significantly, which may be associated with inhibiting the formation of tumor vessel.