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目的:通过动物实验探讨99 Tcm-HYNIC-Annexin V活体细胞凋亡检测筛选肝癌敏感化疗药物的可行性。方法:20只VX-2荷瘤免随机分为4组,通过治疗剂量的紫杉醇(PAC,n=6),5-氟尿嘧啶(5-FU,n=5),表柔比星(EPI,n=5)分组干预治疗,同时建立对照组(C,n=4)。分别予化疗前2h、化疗后46h静脉注射99 Tcm-HYN-IC-Annexin V,2h后再以单光子发射计算体层摄影(SPECT)成像,选取肿瘤部位放射性计数(T)及腿部非肿瘤区部位放射性计数(NT),计算其比值(T/NT),进行半定量分析,确定敏感化疗药物。取肿瘤组织,分别予免疫组化(Tunel)和流式细胞仪(FCM)分析组织细胞凋亡率,放射性核素聚集显像与肿瘤细胞凋亡进行相关性分析。实验数据以SPSS 17.0软件分析。结果:99 Tcm-HY-NIC-Annexin V放射显像显示,化疗前各组肿瘤部位均无明显显像(F=0.025,P=0.980),放射性核素分布主要聚集在肾脏及膀胱区。药物干预48h后,对照组仍无显像,而药物干预组显像清楚,放射性计数显著高于对照组(F=16.52,P<0.001)及化疗前(P<0.01)。药物干预组中,PAC组肿瘤组织放射性显像显著高于5-FU组和EPI组(P值分别为0.042和0.003),而5-FU组与EPI组间放射性显像差异无统计学意义(P=0.224)9。9 Tcm-HYNIC-Annexin V活体细胞凋亡检测结果与免疫组化及流式细胞检测结果一致(R2=0.537 7,P=0.002;R2=0.695 4,P<0.001)。结论9:9 Tcm-HYNIC-Annexin V核素显像检测能够反映活体组织细胞凋亡情况,该方法能够在活体状态下筛选敏感化疗药物。
OBJECTIVE: To investigate the feasibility of screening 99 Tcm-HYNIC-Annexin V apoptosis in vivo to screen chemosensitive drugs for hepatocellular carcinoma through animal experiments. Methods: Twenty tumor-free VX-2 tumors were randomly divided into 4 groups: treated with paclitaxel (PAC, n = 6), 5-fluorouracil = 5) group intervention treatment, while establishing the control group (C, n = 4). Two hours before chemotherapy and 46 hours after chemotherapy, intravenous injection of 99 Tcm-HYN-IC-Annexin V was given to the rats for 2 hours. SPECT imaging was performed with single photon emission computed tomography. Radioactive count (T) and non-tumor District area radioactivity count (NT), calculate the ratio (T / NT), semi-quantitative analysis to determine the sensitive chemotherapeutic drugs. The tumor tissues were taken out and analyzed by Tunel and flow cytometry (FCM) respectively to analyze the correlation between apoptosis rate and radionuclide accumulation imaging and tumor cell apoptosis. The experimental data were analyzed by SPSS 17.0 software. Results: 99 Tcm-HY-NIC-Annexin V radio-imaging showed no significant imaging of the tumor sites (F = 0.025, P = 0.980) before radiotherapy. Radionuclide distribution mainly concentrated in the kidney and bladder area. After 48 hours of drug intervention, there was still no imaging in the control group, while the drug intervention group was clear and the radioactivity count was significantly higher than that in the control group (F = 16.52, P <0.001) and before chemotherapy (P <0.01). In the intervention group, the radioactive imaging of tumor tissue in PAC group was significantly higher than that in 5-FU group and EPI group (P values were 0.042 and 0.003, respectively), but there was no significant difference between 5-FU group and EPI group P = 0.224) .9.9 The results of Tcm-HYNIC-Annexin V in vivo apoptosis assay were consistent with the results of immunohistochemistry and flow cytometry (R2 = 0.537 7, P = 0.002; R2 = 0.695 4, P <0.001). Conclusion 9: 9 Tcm-HYNIC-Annexin V radionuclide imaging detection can reflect the apoptosis of living tissue cells, this method can screen sensitive chemotherapeutic drugs in vivo.