合用阿伐斯汀增强索拉非尼对肝癌生长转移的抑制作用

Effects of avastin combined with sorafenib in a highly metastatic murine model of human hepatocellular carcinoma

目的 观察合用阿伐斯汀（avastin）与索拉非尼（sorafenib）对人肝癌裸鼠模型的抑制效果.方法 采用人肝癌裸鼠原位模型LCI-D20,分为对照组、serafenib单用组（30mg/kg灌胃,1 d1次）、sorafenib与avastin合用组、avastin单用组（5 mg/kg腹腔注射,1周2次）,治疗4周后观察肿瘤体积、肺转移、血浆血管内皮生长因子（VEGF）,定量比较肿瘤微血管密度（MVD）和肿瘤血管内皮细胞凋亡指数,观察各组荷瘤鼠生存期.结果 对照组、sorafenib治疗组、mrafemb与avastin合用组、avastin治疗组肿瘤体积分别为（5.70±0.17）、（1.10±0.18）、（0.60±0.12）和（2.10±0.28）mm^3;肺转移灶数目分别为（191±23）、（98±18）、（31±19）和（98±20）个;血浆VEGF分别为（1689±612）、（3762±1195）、（1844±746）和（420±161）ng/L;中位生存期分别为70、87、112、80 d.4组肿瘤微血管密度分别为（3.77±0.44）%、（1.28±0.15）%、（0.56±0.08）%、（1.32±0.18）%;肿瘤血管内皮细胞凋亡指数分别为（12.6±1.8）%、（32.6±8.7）%、（54.3±11.9）%、（26.8±6.5）%;与sorafenib治疗组比较,avastin与sorafenib合用明显抑制血浆VEGF（P〈0.05）、降低肿瘤体积（P〈0.05）、抑制肺转移灶数目（P〈0.01）,延长荷瘤鼠生存期（P〈0.05）,降低肿瘤微血管密度（P〈0.05）和促进肿瘤血管内皮细胞凋亡（P〈0.05）.结论 sorafenib在肝癌裸鼠模型中上调血浆VEGF导致耐药,avastin下调VEGF,通过促进肿瘤血管内皮细胞凋亡和降低肿瘤微血管密度,进一步增强sorafenib对肝癌生长和转移的抑制作用,并延长荷瘤鼠生存期.

Objective To investigate the effect of combination of sorafenib and avastin,a vascular endothelial cell growth factor （VEGF） neutralizing antibody on tumor growth, lung metastasis and survival of tumor-bear nude mice in a highly metastatic xenograft murine model of human hepatocellular carcinoma （HCC）. Methods Xenograft of a highly metastatic human HCC tumor （LCI-D20） was used to evaluate survival, primary tumor growth and lung metastasis after treatment with avastin alone or in combination with sorafenib. Tumor angiogenesis was identified by immunohistochemistry staining for CD31 and calculated as microvessel density （MVD）. Apoptosis of tumor endothelial cells were determined by double immanofluor-esence staining for CD31 and the terminal deoxynucleotidyl-transferase mediated dUTP nick-end labeling assay （TUNEL）. Results Tumor volumes were （5.70±0.17 ）, （1.10±0.18）, （0.60±0.12） and （2.10±0.28） mm^3 in control,serafenib,sorafenib plus avastin and avastin group. Lung metastasis was （191±23）, （98±18）, （31±19） and （98±20） in four groups, and median survival were 70,87,112 and 80 days,respectively. MVD were （3.77±0.44）%, （1.28±0.15）%, （0.56±0.08）%, （1.32± 0.18）% ,and indexes for apoptotic endothelial cells were （12.6±1.8）%, （32.6±8.7）%, （54.3± 11.9） %, （26.8±6.5） % respectively in four groups. Combination of avastin and serafenib significantly reduced plasma VEGF （P〈0.05） ,decreased tumor volume （P〈0.05） ,inhibited numbers of lung metas-tasis （P〈0.01）, and prolonged survival compared to mice treated with serafenib alone （P〈0.05）. Immu-nofluoresence staining shows reduced MVD （P〈0.05） and increased apoptosis of tumor endothelial cells （P〈0.05） by combination of avastin and sorafenib. Conclusion VEGF may contribute to resistance of sorafenib, avastin significantly inhibited tumor angiogenesis by inducing apoptosis of tumor endothelial cells. The study implies that it is promising to combine avastin with sorafenib for patients with advanced hepatocellular carcinoma.