【作者】 陈长广；

【导师】 倪才方；

【作者基本信息】 苏州大学 ， 影像医学与核医学， 2007， 硕士

【摘要】 目的:通过动物实验研究射频消融联合肝动脉栓塞治疗肝癌后残余肿瘤生物学行为的变化,初步探讨联合治疗的机制,评价其治疗效果。方法:健康清洁级新西兰大白兔45只在CT引导下行VX2瘤块种植法建立成肿瘤直径2.5cm～2.7cm大小的肝癌动物模型。建模成功后随机分成对照组(假性治疗组)、单纯射频治疗组和肝动脉碘油栓塞7天后射频治疗组(单极射频针裸露端长均为2cm),共3大组。每组分别于治疗结束后1天、4天、7天处死5只瘤兔。切取肿瘤中心组织及边缘组织为检测标本,全部组织经10%Formalin固定,常规石蜡包埋,4μm连续切片,对取材标本行HE染色观察治疗后病理形态学变化。肿瘤边缘组织行免疫组织化学方法检测残余肿瘤细胞凋亡、增殖细胞核抗原、血管内皮生长因子及微血管密度的表达。所有数据均用统计软件包SPSS13.0处理。计量资料用X±S表示,以P<0.05有统计学差异。结果:1.TUNEL法及Caspase-3检测细胞凋亡结果显示:单纯射频及联合治疗组凝固性坏死区周边的肿瘤细胞凋亡指数(AI)明显高于对照组(P<0.01),以射频后1天最为明显,后逐渐减少,7天后虽明显减少但仍高于术前水平。且联合治疗组各时间点残余肿瘤细胞凋亡指数也高于单纯射频组(P<0.01)。2.单纯射频及联合治疗组残余肿瘤细胞增殖指数(PI)明显低于对照组(P<0.01),且联合治疗组残余肿瘤细胞增殖指数(PI)也低于单纯射频组(P<0.05)。3.单纯射频及联合治疗组VEGF低于对照组(P<0.05)。联合治疗组VEGF绝对值略高于单纯射频组,但差异无统计学意义(P>0.05)。4.单纯射频及联合治疗组残余肿瘤微血管密度(MVD)明显低于对照组(P<0.01)。联合治疗组残余肿瘤微血管密度(MVD)绝对值略高于单纯射频组,但差异无统计学意义(P>0.05)。5.残余肿瘤细胞VEGF与MVD表达呈正相关(P<0.01)。结论:1.单纯射频和射频消融联合肝动脉栓塞治疗兔肝VX2肿瘤一周内均能诱导残余肿瘤细胞凋亡,同时抑制肿瘤细胞增殖。2.联合治疗与单纯射频治疗相比更能促进残余肿瘤细胞凋亡,抑制肿瘤细胞增殖。3.单纯射频和联合治疗一周内均能抑制肿瘤血管生成,且联合治疗在抑制肿瘤血管生成作用方面与单纯射频治疗作用相当。4.联合治疗后残余肿瘤内仍有VEGF表达,仍存在肿瘤血管生成。临床上如联合应用抗肿瘤血管生成药物可能会取得更好的疗效。更多还原

【Abstract】 Objectives: To study the biological behavior of remnant tumor of hepatocellular carcinoma (HCC) after radiofrequency ablation (RFA) combined with transcatheter arterial embolization (TAE); to investigate the mechanism of the combined therapy; and to assess the therapeutic efficacy of it.Methods: Forty-five New Zealand white rabbits were implanted percutaneously with VX2 tumor cells in the left lobe of the liver under CT guidance successfully. Three weeks later, there was a focal tumor (φ2.5cm-2.7cm) in the liver of each rabbit. Then they were divided into 3 groups at random: control group (pseudo-therapy group), RFA (single electrode 2.0cm exposure) alone group and 7 days after TAE combined with RFA (single electrode 2.0cm exposure) sequentially group. Each group was divided into 3 subgroups respectively according to the killing time: 1 day, 4 days and 7 days after treatment. All the 45 rabbit-models were sacrificed after treatment and their tissues,including peripherial tumor and central tissue of the ablation area,were sectioned, embedded in paraffin, fixed in 10% Formalinand and spitted constantly with 4μm–section. They were examined pathologically by HE dying and immunohistochemical methods (Apoptosis, CD31, VEGF, PCNA). The results of immunohistochemistry were compared. All the data were analysed by SPSS 13.0 software. Quantitative data were showed by x±s, P<0.05 was defined as threshold for statistical significance .Results: 1. Quantification assay of apoptosis measured by TUNEL and Caspase-3 staining methods indicated that apoptosis index (AI) in the margin of the tumor after RFA alone and combined therapy were both higher than that of control group significantly (P<0.01), they reached their peaks at 1 day, and decreased gradually along with the time to a low level at 7 days after treatment , but were still higher than that of pre-treatment . And AI of each subgroup after combined therapy were higher than that of RFA alone therapy significantly (P<0.01) . 2. Proliferation index (PI) of survival tumor cells after RFA alone and combined therapy were both lower than control group significantly (P<0.01) , and PI of each subgroup after combined therapy were lower than that of RFA alone therapy (P<0.05) . 3. Vascular endothelial grow factors (VEGF) of survival tumor cells after RFA alone and combined therapy were both lower than control group significantly (P<0.05). VEGF of each subgroup after combined therapy were higher than VEGF of RFA alone therapy slightly , but there was no statistical significance between them (P<0.05). 4. Microvessel density (MVD) of survival tumor cells after RFA alone and combined therapy were both lower than control group significantly (P<0.01) . MVD of each subgroup after combined therapy were higher than MVD of RFA alone therapy slightly, but there was no statistical significance between them (P<0.05) . 5. There was a positive correlation between VEGF and MVD in the combined treatment groups (P<0.01) .Conclusions: 1. Both RFA alone and RFA combined with TAE therapy can induce the apoptosis and inhibit the cell proliferation in 1 week after treatment. 2. Compared with RFA alone , combined therapy can induce more cells to apoptosis and inhibit more cells proliferation , and can get better therapeutic efficacy . 3. Both RFA alone and RFA combined with TAE therapy can effectively inhibit tumor angiogenesis in 1 week after treatment. As for the efficacy of it, combined therapy is parallel with RFA alone. 4. Duing to tumor angiogenesis existed, combined therapy plus anti-angiogenesis drugs may obtain better therapeutic efficacy.更多还原