【作者】 牛陵川；

【导师】 王智彪；

【作者基本信息】 重庆医科大学 ， 生物医学工程， 2010， 博士

【摘要】 目的1.建立聚焦超声消融后家兔残留乳腺肿瘤模型;2.综合评价聚焦超声消融后即刻辐照靶区肿瘤组织和残留肿瘤组织的病理学变化情况;3.研究聚焦超声消融后残留乳腺肿瘤组织增殖能力变化的规律;4.研究聚焦超声消融后残留乳腺肿瘤对机体的影响。方法1.利用组织块包埋法先建立兔VX2种植性乳腺肿瘤模型,当肿瘤直径约10mm～15mm时接受FU消融治疗。在US引导和监控下消融大部分肿瘤组织,通过热电偶数字显示测温计控制治疗区旁肿瘤组织的温度,避免完全消融肿瘤以建立聚焦超声消融后家兔残留乳腺肿瘤模型。2.FU消融后即刻,对US辐照靶区肿瘤组织和残留肿瘤组织进行病理学检查:应用HE染色光镜观察常规组织学改变;应用电镜观察肿瘤细胞超微结构的改变;应用酶组织化学染色判断肿瘤细胞功能活性。3.FU消融后的6个不同时间点(即刻、3d、7d、14d、21d和28d),应用免疫组织化学方法检测增值细胞核抗原(PCNA)和Ki-67表达的变化情况。4.通过尸解观察家兔在不同时间点肿瘤的转移情况以及家兔自身的自然生存时间。结果1.FU消融后即刻,辐照靶区肿瘤组织的HE染色显示瘤细胞形稍肿胀,胞内出现大小不一的空泡,细胞核形态未见明显改变;电镜显示肿瘤细胞大体轮廓仍然存在,细胞膜连续性完全丧失,细胞器结构破坏或消失,胞浆内甚至胞核内出现大小不等的空泡,核膜断裂,染色质固缩、碎裂呈团块状,呈典型凝固性坏死表现;酶组织化学染色显示肿瘤细胞SDH和ACP无表达;免疫组织化学显示PCNA和Ki-67亦呈阴性表达。2.FU消融后即刻,残留肿瘤组织的HE染色显示瘤细胞的细胞形态和细胞核形态均未见明显改变;电镜显示瘤细胞明显水肿、细胞器结构肿胀,呈热损伤表现;酶组织化学染色显示肿瘤细胞SDH和ACP活性降低;PCNA和Ki-67表达明显下降。3.FU消融后,残留肿瘤组织的PCNA和Ki-67表达呈一过性降低,14d后逐渐回升,21d恢复至消融前水平,消融后28d也未见其表达明显增高。4.残留肿瘤家兔最早出现肿瘤转移的时间是消融后21d,至消融后28d,其淋巴结总转移率为13.3%(4/30),脏器总转移率为6.7%(2/30);未接受FU消融处理的家兔最早出现肿瘤转移的时间是消融后7d,至消融后28d,其淋巴结总转移率为50%(9/18),脏器总转移率为38.9%(7/18)。5.残留肿瘤家兔的自然生存时间为64.8 d±10.4d(45d～80d),较未接受FU消融处理的家兔生存时间45.6 d±11.0d(31d～59d),延长了42%。结论1.当肿瘤直径约10mm～15mm时,为家兔VX2种植性乳腺肿瘤模型进行FU消融处理的最佳时期。2.FU消融后即刻,HE染色光镜下无法判断肿瘤细胞的存活情况,但电镜和酶组织化学染色(SDH和ACP)检查均能有效证实FU消融靶区肿瘤组织的瘤细胞已经死亡。3.经电镜和酶组织化学染色检查证实,通过控制探针处肿瘤组织的温度(最高温升不超过50℃,且持续时间不超过10s),在确保靶区消融效果的前提下建立残留乳腺肿瘤模型是成功的。4.FU未能一次完全性消融乳腺肿瘤,在短时间内也能有效抑制残留肿瘤的生长和转移,延长机体的生存时间。更多还原

【Abstract】 Objective1. To establish the rabbit VX2 remnant breast tumor model with focused ultrasound ablation.2. To evaluate the pathological changes of the target tumor tissue and remnant tumor tissue immediately after focused ultrasound ablation.3. To observe the changes in proliferative capability of remnant breast tumor implanted in rabbits after focused ultrasound ablation.4. To study the effects of the remnant breast tumor on the body.Methods1. First,to establish rabbit VX2 implanted breast tumor model of using tissue mass embedding method;Then,FU was applied to the tumors and the remnant tumors were obtained by controlling temperature of the tumor tissue around the thermal sensor.2.The ablated and residual tumor tissue were both evaluated immediately after FU ablation by HE staining,electron microscopy and enzyme histochemistry staining (SDH and ACP) to confirm if the remnant tumor model has been established.3. The expression of PCNA and Ki-67 was determined by immunohistochemistry at different time points after FU ablation(immediately,3d,7d,14d,21d and 28d).4.The organ and lymph node metastases,and the natural survival time were observed after FU ablation.Results1. In the FU ablated tumor tissue immediately after FU ablation,the outline of cells remained,many vacuolat-like structures appeared in cells,and there were no significant changes in nucleus under light microscope;and electron microscopy showed that the outline of cells remained,but the continuity of cell membrane was lost,organelle structure was damaged or disappeared,nuclear membrane was ruptured,different sizes of vacuoles in cells,nuclear membrane ruptured,chromatin condensed and fragmented into massive structures;and the activity of SDH and ACP was completely inactivated;and the expression of PCNA and Ki-67 was negative.2. In the residual tumor tissue immediately after FU ablation,there was no significant change in cell morphology and nuclear morphology under light microscope;and electron microscopy showed the thermal injury performance that includes the edema of tumor cells and swelling structure of organelles;the activity of SDH and ACP was decreased;and the expression of PCNA and Ki-67 was significantly decreased.3. The expression of PCNA and Ki-67 in residual tumor cells were decreased after FU ablation for a short time,it was gradually recovered at day 14,and restored to the level before ablation at day 21,and without further significantly increase at day 28.4. In the remnant tumor group,the earliest metastasis time was at day 21 after ablation,and the total lymph node metastasis rate was 13.3% and total visceral metastasis rate was 6.7% on day 28 after ablation. In the control group,the earliest metastasis time was at day 7 after ablation,and the total lymph node metastasis rate was 50% and total visceral metastasis rate was 38.9% on day 28 after ablation.5. The mean natural survival time of rabbits in the remnant tumor group was 64.8d and certainly prolonged by 42%.Conclusion1. It is considered that the rabbit VX2 implanted breast tumor model was successfully built and the best time for FU ablation is when the tumor is 10mm～15mm in diameter.2. Although HE staining under light microscope can not determine the survival condition of VX2 tumor cells,however,the electron microscopy and enzyme histochemical staining both confirmed that the tumor cells are dead.3. By controlling the temperature of tumor tissue around thermal senser(the temperature was under 50℃and last less than 10s),the rabbit VX2 residual tumor model was successfully established.4. Though FU ablation was not completely eradicate tumor,the growth and metastasis of the rabbit VX2 residual breast tumors were effectively inhibitted for a short period time,and it can prolong the survival time of the subjects.更多还原