抗骨桥蛋白抗体抑制HCCLM3裸鼠移植瘤的生长并增强其对放疗敏感度的实验

目的 探讨抗骨桥蛋白抗体对人肝癌HCCLM3细胞裸鼠移植瘤生长、转移和血管生成的抑制作用及对放疗的增敏作用.方法 74只裸鼠右侧腹皮下接种肝癌细胞株HCCLM3,次日随机取50只裸鼠随机分为5组,每组10只:A组尾静脉注射0.9％氯化钠溶液,B组尾静脉注射非特异性抗体,C、D、E组分别尾静脉注射抗OPN抗体5、10、20 mg/kg,每周2次.每两天观察肿瘤体积变化,第10周末处死裸鼠,收集肿瘤组织、肺组织进行病理组织学检查,用免疫组织化学法测肿瘤微血管密度.其余24只裸鼠随机分为4组:F组给予尾静脉注射0.9％氯化钠溶液、G组给予荷瘤鼠肿瘤局部以6 MeV电子线照射、H组给予尾静脉注射抗OPN抗体20 mg/kg、Ⅰ组给予尾静脉注射抗OPN抗体20 mg/kg+肿瘤局部6 MeV电子线照射.观察肿瘤体积变化,比较各组的肿瘤体积、抑瘤率,利用增敏系数(enhancement factor,EF)评价抗OPN抗体的增敏作用.结果 与A、B对照组相比,抗OPN抗体均能明显抑制肿瘤的生长(P＜0.01)、降低肿瘤组织中血管密度(P＜0.05)、减少肺转移(P＜0.05),其中以20 mg/kg剂量组明显.在放疗的实验中,抗体联合放疗组抑瘤率明显高于单纯放疗组和抗体组(P均＜0.01).EF＞1,提示抗OPN抗体对移植瘤有放疗增敏作用.结论 抗OPN抗体明显抑制了肿瘤的生长、转移、血管生成并能提高肝癌对放疗的敏感度,它有望成为治疗肝癌的又一武器.

Experiment of Anti-OPN Antibody Inhibits Growth and Increases Radiosensitivity of Tumors in NudeMice Implanted with HCCLM3

Objective
To study the inhibiting effect of anti-OPN antibodies on tumor growth,metastasis,angiogenesis
of HCCLM3 tumor in nude mice and its enhancement effect on radiotherapy.MethodsHCCLM3 wereinoculated subcutaneously into the right flank of 74 Balb/c nude mice.Next day after inoculation,50mice were randomized into five groups（10 mice per group）：Group A (treatment with normalsaline),Group B (treated with non-specific antibody),Group C,D,E (treated with 5 mg/kg,10 mg/kg,20mg/kg of the anti-OPN antibodies,respectively).The treatments were repeated twice per week and tumorvolume was measured every other day.
At the end of the 10th week,nude mice killed,tumors and lungs of the mice were collected.Microvessel density (MVD) of tumor tissue was measured by IHC.The other 24 nude mice bearing HCCtumors were randomly divided into four groups：Group F (treated with normal Saline),Group G (treatedwith 6 MeV electron ray on the tumor),Group H (treated with 20 mg/kg anti-OPN antibody),Group I(treated with 20 mg/kg anti-OPN antibody and 6 MeV electron ray on the tumor).The tumor volume wasmeasured every day and the enhancement factor (EF) was used to evaluate the radiationenhancement.Results
Anti-OPN antibody had significant inhibiting effect on the tumor growth (P<0.01),as well as reducedmetastasis (P<0.05) and angiogenesis (P<0.05) when comparing with control groups A,B.
And among all of them, 20 mg/kg was the most remarkable dose.
The inhibition rate of partial tumor growth in combination group I (41.67%),was higher than that ofanti-OPN antibody group H （11.86%,P<0.01） and radiation group G(24.04%)（P<0.01).
EF>1,which meant anti-OPN antibody had radiotherapy enhancement effect on implented tumor.
ConclusionThe anti-OPN antibody can inhibit the tumor growth,metastasis,and angiogenesis and promotethe radiotherapy enhancement of liver cancer,which would be another weapon for the HCC.