单次化疗后早期肿瘤内^(99)Tc^m-rh-Annexin Ⅴ分布与VEGF、Survivin表达的相关性研究

Distribution of ^(99)Tc^m-rh-Annexin Ⅴ and its relationship with expression of VEGF and Survivin in tumor after a single dose of chemotherapy

目的:探讨单次化疗前、后肿瘤组织内99Tcm-重组膜联蛋白V(rh-Annexin V)的分布与血管内皮生长因子(VEGF)表达及Survivin蛋白表达的相关性及其潜在临床应用价值。方法:采用99Tcm直接标记法标记rh-Annexin V,作为核素凋亡显像示踪剂。将小鼠肝癌细胞Hca-F25接种于小鼠右腋窝皮下;随机分为A(n=9,对照组)、B(n=10,化疗组)两组;肿瘤生长至直径约1cm时,B组一次性接受环磷酰胺腹腔内注射,剂量为150mg/kg;20h后两组同时由鼠尾静脉注入99Tcm-rh-Annexin V[3.7～7.4MBq/(0.5μg·0.1ml)];4h后行SPECT显像并处死、取材,应用井型闪烁计数器检测荷瘤小鼠肿瘤组织内示踪剂分布,以每克组织百分注射剂量率(%ID/g)表示。采用免疫组化SP法检测标本中VEGF及Survivin蛋白表达。结果:B组肿瘤组织内99Tcm-rh-Annexin V分布明显多于A组,而Survivin蛋白表达A组明显高于B组,差异均有显著统计学意义(P<0.01);VEGF表达在A、B组中无显著差异(F=0.072,P>0.05)。相关性研究表明,A组肿瘤组织内,VEGF表达与99Tcm-rh-AnnexinV分布呈明显负相关(n=9,r=-0.713,Y=-8.139X+4.395,F=7.240,P=0.031),与Survivin蛋白表达呈明显正相关(n=9,r=0.776,Y=0.848X-1.888,F=10.573,P=0.014);B组肿瘤组织内,VEGF表达与99Tcm-Annexin V分布及Survivin蛋白表达均无明确相关性。肿瘤组织内Sur-vivin蛋白表达与99Tcm-Annexin V分布在A、B组内呈明显负相关。结论:化疗后早期,肿瘤组织内99Tcm-rh-Annexin V分布较VEGF表达更能真实地反映肿瘤细胞的凋亡状态;VEGF抑制肿瘤细胞凋亡的特性可能需要高表达的Survivin蛋白所介导。

Objective： To investigate the distribution of 99Tcm-rh-Annexin V and its relationship with expression of VEGF and Survivin after a single dose of chemotherapy. Methods： Eight days after being inoculated with allogenic hepatoma cells （Hca-F25） into the subcutaneous tissue of the right axillary fossa, the mice （purebred 615） were randomly divided into two groups （group A was the control group, n=9; group B was the treated group, n=10）. Group B received a single dose of chemotherapy intraperitoneally （cyclophosphamide, 150mg/kg）. 99Tcm-rh-annexin V （3.7-7.4MBq/0.5μg.per mouse） were injected intravenously 20h later. Four hours after 99Tcm-rh-Annexin V injection, the animals were imaged and sacrificed, the tumor samples were weighed and the radioactivity was determined with a well-type scintillation counter. The accumulation of 99Tcm- rh-annexin V in tumor was expressed as the %ID/g （percentage activity of injection dose per gram of tissue）, and the expression of VEGF and Survivin in tumor were determined with immunohistochemical methods. Results： Single dose of chemotherapy significantly increased the tumor uptake of 99Tcm-rh-Annexin V （P〈0.01）, but the expression of Survivin was significantly high in group A than in group B（P〈0.01）. There were no statistical significance between group A and group B in the expression of VEGF. The expression of VEGF correlated negatively with the distribution of 99Tcm-rh-Annexin V and positively with expression of Survivin in group A, but it had no relationship with these in group B. The distribution of 99Tcm-rh-Annexin V correlated negatively with the expression of Survivin in both groups. Conclusions： The distribution of 99Tcm-rh-Annexin V in tumor could reflect the degree of apoptosis early after a single dose of chemotherapy more truly than the expression of VEGF, and maybe the anti-apoptosis function expression of VEGF need high level expression of Survivin in tumor.