兔肝Vx-2移植癌改良模型的建立

 目的 培养、建立稳定的兔肝Vx|2移植癌模型，探讨不同植瘤方式的成功率。方法 60只新西兰白兔随机分3组，每组20只。 分别以不同的方式将Vx|2瘤细胞植入兔的肝脏。观察：1.不同方式植瘤的成活率；2.B超测定肝内肿瘤7d、10d、14d、17d、21d时的大小，并计算肿瘤生长率；3.大体及镜下(光镜和电镜)瘤组织形态特征。结果 1.三组植瘤成活率分别为7/20、10/20、19/20，改良组植瘤成活率最高（P<0.05)，瘤体呈指数性生长；2.病理学及CT表明该瘤在肝组织中呈浸润式生长，其性状与移植于兔其它部位的Vx|2鳞状细胞癌特征相似。结论 成功建立了兔肝Vx|2移植癌模型，瘤组织块种植方式成功率明显高于其它两组方式，为肝癌介入治疗的基础及临床研究，提供了成熟的大型实验动物模型。     

兔鼻咽VX2移植癌模型的建立及其生长转移特性

目的:建立兔鼻咽VX2移植癌动物模型,观察肿瘤局部生长浸润、淋巴结和远处转移的特性及在18F-FDG PET-CT、MRI图像上的表现.方法:将VX2瘤细胞株荷瘤新西兰白兔麻醉后,解剖将肿瘤剥离,制成1×107～2x107/mL细胞悬液,注入免腹股沟,形成传代兔传代.在CT引导下,将瘤细胞悬液注入兔鼻咽部,建立兔鼻咽VX2移植癌动物模型后.观察兔症状及肿瘤生长、淋巴结和远处转移特征及影像学表现.结果:41只兔在鼻咽种植瘤细胞2～4周内.37只在鼻咽部形成实体瘤,成功率90.2%(37/41),7只意外死亡,30只进入研究.鼻咽部肿瘤成瘤后均增长迅速.向周围浸润性生长,常见的周围浸润部位有咽旁间隙、鼻腔、口咽、颅底、斜坡、颞下窝、海绵窦等.病理检查符合低分化鳞状细胞癌表现.颈部淋巴结转移率93.3%(28/30),双颈淋巴结转移率43.3%(13/30).2例出现肺多发转移6.7%(2/30).30只兔鼻咽结构、周围组织器官及鼻咽肿物、淋巴结转移灶、远处转移灶在18F-FDGPET-CT图像上均能清晰显示.结论:兔鼻咽VX2移植癌动物模型建立成功,且成功率高.鼻咽部肿瘤成瘤后迅速向周围浸润性生长,颈部淋巴结转移率高,在18F-FDGPET-CT、MRI图像上能清晰显示鼻咽腔内外结构以及与周围组织的关系,提供影像证据,是进行人类鼻咽癌实验研究的理想动物模型.     

肿瘤细胞悬液注射法制作兔食管癌移植瘤模型

目的:利用肿瘤细胞悬液注射法建立稳定的兔食管癌移植瘤模型,观察其钡餐、MRI及病理表现.方法:冻存的VX2肿瘤细胞经兔体内接种成瘤后制成细胞悬液,手术暴露食管,直视下将肿瘤细胞悬液接种于兔食管壁,制作兔食管癌移植瘤模型,2周后经钡餐和MRI检测其影像学表现,处死兔子取标本行病理学检测.结果:食管内注射肿瘤细胞悬液2周后,兔子出现吞咽困难的症状,食管钡餐显示明显的充盈缺损,缺损内径平均约0.518cm;MRI显示兔食管壁内孤立性结节状肿瘤,病灶在T1W1上呈略低信号或等信号,在T2W1上呈高信号;瘤体和淋巴结标本病理切片镜下观察,证实成瘤及出现淋巴结转移;模型建立成功率为100%.结论:使用肿瘤细胞悬液注射法可成功建立兔VX2食管癌动物模型.     

人卵巢癌裸鼠移植瘤和腹水瘤模型的建立及形态学观察

目的 建立人卵巢癌裸鼠移植模型,为研究人卵巢癌发病机制、实验治疗提供工具。方法 采用组织学完整 的人卵巢癌实体瘤和腹水瘤标本植入裸鼠腹腔和皮下,观察成瘤生长和转移以及形态学特征（光镜、电和染色体）。结果 10例卵巢癌标本2例实体瘤、1例腹水瘤移植成功,实体瘤传至第六代,腹水瘤传至第十八代,传代移植成功率100％,移植瘤病理学和电镜观察、染色体核型分析、肿瘤标记物检查均与人体原发肿瘤一致。结论 本研究建立的人卵巢癌实体瘤和腹水瘤裸鼠移植模型与临床患者相似,为进一步研究人体卵巢癌发病和治疗提供了理想的实验动物模型。     

荷人肺癌小鼠皮下移植瘤模型的建立及其生物学特性初探

背景与目的 为了更好地研究肺癌的治疗方法,建立起可靠的动物评价模型迫在眉睫.本研究的目的是研究建立肺癌原代组织块小鼠移植瘤模型的成熟方法,观察移植瘤的肿瘤生物学特性,在建模方法及基本特征等方面来证明该肿瘤模型的合理性和科学性,以期为肿瘤研究提供更有效的实验动物模型.方法 取人新鲜完整肺癌组织块移植于NOD/SCID小鼠右侧前肢肩背部皮下,或经皮肺穿刺取得肿瘤小块移植于BALB/c裸小鼠肾包膜下.待皮下肿瘤增大,将其切下传代于裸小鼠右侧前肢肩背部皮下.观察移植瘤生物学特性,并取肿瘤行常规病理切片及CEA、细胞角蛋白、Ki67免疫组化检测,将原代肿瘤和移植瘤进行EGFR 18-21外显子和K-Ras 12,59外显子基因检测,采用流式细胞仪检测移植瘤细胞的细胞周期.结果 本研究进行了11例肺癌组织的NOD/SCID小鼠和裸鼠建模,建成3例可多次成功传代的腺癌、小细胞肺癌和鳞癌模型.传代移植成功率高.荷瘤小鼠生长情况良好,生存期长.各代移植瘤模型的组织病理学及免疫组化表型、EGFR和K-Ras基因检测均与来源肺癌组织相一致.移植瘤细胞周期中S期延长,提示瘤细胞有增殖活性.结论 本研究在国内首次利用新鲜的完整肺癌组织建成了荷肺癌NOD/SCID小鼠及裸鼠模型,并传代移植于裸鼠,建模成功率为27%.移植瘤较好地保留了人原发肺癌的恶性特征及组织病理学、生物学特性,是一种接近人体的肺癌模型,可为肺癌研究提供良好的实验平台.     

兔VX2食管癌模型的建立及生物学特性观察

目的:建立兔VX2食管癌移植瘤模型,探讨其成瘤率、生长特性及转移情况,为进一步研究食管癌及寻找有效治疗方法提供平台.方法:将兔VX2肿瘤剪碎至1 mm3左右的组织块,通过外科手术将肿瘤块植入腹段食管黏膜下层,于接种后第1、2及3周分别行食管造影和CT扫描检查,观察肿瘤生长情况.3周后,处死兔子取标本行病理学检测.结果:12只兔子除2只死于手术并发症外(胃穿孔及误缝食管所致的梗阻),其余10只接种后1周,未见肿瘤生长;2周后食管造影显示食管下段充盈缺损.CT扫描示食管下段走行区局限性管壁增厚,有明显软组织肿块影,平均最大径为1.04 cm (0.7～1.5 cm);3周后兔子消瘦明显,食管造影示食管下段黏膜破坏,充盈缺损明显,4只兔子出现食管穿孔.CT扫描示食管下段软组织肿块影明显增大,平均最大径为2.28 cm (1.7～4.3 cm).瘤体及淋巴结病理检查证实,成瘤及淋巴结转移成功率为100%.结论:采用兔VX2瘤块移植法可成功建立食管癌移植模型,且具有周期短,成瘤率高,生长迅速等特点.     

4种人肾细胞癌原位移植裸鼠模型建立方法的比较

背景与目的:肾细胞癌是最常见的肾脏恶性肿瘤,起病隐匿,恶性程度高.研究旨在建立成功率高、稳定的人肾细胞癌原位动物造模方法.方法:采用人肾细胞癌细胞(786-0、ACHN)进行细胞悬液原位注射法、皮下成瘤后引瘤原代细胞悬液原位注射法、皮下成瘤后引瘤组织块原位移植肾包膜下法及皮下成瘤后引瘤组织块原位移植肾周筋膜内法建立原位移植裸鼠模型,采用PET/CT、H-E染色、免疫组织化学染色及血清生化检测等手段观察成瘤率及肿瘤生长情况,评估4种方法的建模效果.结果:人肾细胞癌皮下移植瘤裸鼠模型,ACHN组成瘤率(90%)高于786-0组(30%);人肾细胞癌原位移植裸鼠模型,786-0原位注射组、ACHN原位注射组、ACHN皮下移植瘤引瘤后原代细胞原位注射组、ACHN组织块肾包膜下包埋组和ACHN组织块肾周筋膜内包埋组的成瘤率分别为33%、80%、90%、100%和20%,其中以ACHN皮下成瘤后引瘤组织块原位移植肾包膜下法成瘤率最高,影像学及病理组织学检查结果显示符合低分化肾细胞癌.结论:成功建立4种人肾细胞癌原位移植裸鼠模型,其中以ACHN皮下成瘤后引瘤组织块原位移植肾包膜下法成瘤效果最佳,为肾细胞癌发病机制及靶向治疗的进一步研究提供理想的动物模型.     

人胃癌组织块裸鼠原位移植/转移模型的建立

 目的　用肿瘤组织块原位移植 ,建立人胃癌裸小鼠原位移植 /转移模型。方法　以人胃低分化腺癌细胞系接种于裸小鼠皮下 ,形成稳定传代的皮下移植瘤 ,再取该肿瘤组织块原位移植于裸鼠胃壁 ,观察移植肿瘤的生长状况、移植成功率和自发转移的发生率。结果　原位移植成功率 (成瘤率 )为 1 0 0 %、局部淋巴结转移率 1 0 0 %、远处淋巴结转移率 90 %、肝转移发生率为 75%。荷瘤鼠的中位生存期为 1 4周 ,晚期出现消瘦和全身衰竭。结论　该裸小鼠原位移植 /转移模型的生物学行为与人胃癌自然生长和转移过程相似 ,可作为一种有价值的工具用于胃癌转移机理和抗转移实验治疗的研究。     

兔VX2腹膜转移癌模型的建立及转移特征研究

目的:建立大动物腹膜转移癌模型,鉴定其生物学行为.方法:36只新西兰大白兔分为开腹包埋、开腹穿刺及直接经皮穿刺种植三组(每组12只),每组又分为6只瘤块接种,6只悬液接种,接种VX2肿瘤.观察肿瘤生长状况,通过病理学和影像学检查分析局部区域及远处转移情况.结果:三种方法构建模型的成功率分别是100%(12/12)、91.7%(11/12)和58.3%(7/12),开腹法与经皮穿刺法相比有显著性差异(P＜0.05).接种2周后,形成典型溃疡型胃癌并腹膜转移癌表现,宿主衰竭,4周出现肺转移.病理学检查符合典型VX2组织学特点.结论:开腹包埋与开腹穿刺接种制作VX2兔腹膜转移癌模型简单,实验周期短,接种率高,其病理表现类似人类腹膜转移癌,为腹膜癌治疗的实验研究提供可靠的大型动物模型.     

荷人前列腺癌裸鼠移植瘤模型的建立及其应用研究

目的 应用人前列腺癌细胞株LNCaP接种裸鼠,鼠间移植传代,建立人前列腺癌裸鼠移植瘤模型.方法 观察移植瘤大体形态和组织病理学检查,并应用免疫组织化学方法观察肿瘤组织细胞中前列腺特异膜抗原（PSMA）的表达以及125I标记抗PSMA单抗的荷瘤裸鼠体内放射免疫显像.结果 移植瘤的形态和功能特性与原发肿瘤基本相似,移植瘤的移植成功率为100%;放免显像显示标记抗体能浓聚于肿瘤部位,经尾静脉注射标记抗体后96 h,肿瘤显像清晰,肿瘤组织/非肿瘤组织放射性比值（T/NT）均大于2.8.结论 本动物模型的建立为今后前列腺癌放射免疫治疗的实验研究提供1个理想的模型.     

Magnetic Resonance Evaluation of Transplanted Endometrial Carcinoma and Its Lymph Node Metastasis in Rabbits

Objective： To establish a rabbit model of transplanted endometrial carcinoma with lymph node metastasis and observe its magnetic resonance imaging （MRI） features. Methods： VX2 tumor grafts were orthotopically embedded in the endometrium of rabbits, and 3 weeks after the transplantation, thetumor and its metastasis to the retroperitoneal lymph nodes were examined by MRI, and the signal intensities and size of the lymph nodes were compared with those of normal rabbits. Results： The orthotopic transplantation of the tumor grafts resulted in tumor growth in all the 12 recipient rabbits. The tumors infiltrated the serosa of the uterus and metastasized to the retroperitoneal lymph nodes 3 w after transplantation. MRI demonstrated that the lymph nodes of the tumor-bearing rabbits were larger in size than those of normal control rabbits, but the signal intensity of the lymph nodes was not significantly different between them. Conclusion： This transplanted endometrial carcinoma model is characterized by high success rate and similar tumor metastasis behaviors with human endometrial carcinoma, therefore may serve as a good model for testing the efficacy of contrast agents for MR lymphography.     

快速大容量尾静脉注射法构建循环肝癌细胞肝转移小鼠模型

目的： 通过快速大容量尾静脉注射方法构建循环肝癌细胞肝内复发转移小鼠模型。 方法： 40只C57BL/6J小鼠采用随机数字表法随机分为对照组和实验组（20只/组）,对照组采用传统的缓慢小容量尾静脉注射法（2×10 6个肝癌Hepa1-6细胞/0.2 ml,30 s内注射完毕）,实验组采用改良的快速大容量尾静脉注射法（2×10 6个肝癌Hepa1-6细胞/2 ml,5 s内注射完毕）构建循环肝癌细胞肝内复发转移小鼠模型。4周后摘取肝、肺、肾、脾组织并行H-E染色,大体和镜下观察肝脏、肺脏、脾脏和肾脏的成瘤情况。 结果： 对照组小鼠只在肺脏有转移灶形成,成瘤率为95%（19/20）,肝脏、肾脏、脾脏未见有转移灶形成。实验组小鼠在肝脏和肺脏同时形成转移灶,肺脏成瘤率为94.7%（18/19）,肝脏成瘤率为100%（19/19）,脾脏、肾脏未见转移灶形成。 结论： 快速大容量尾静脉注射法注射循环肝癌细胞构建的小鼠模型可以模拟肝癌根治性切除后残留的循环肝癌细胞导致早期肝癌肝内复发转移的过程。     

三氧化二砷经肝动脉栓塞化疗对兔肝移植瘤的作用

 目的　探讨As2 O3 经肝动脉栓塞化疗对肝移植瘤的作用。方法　采用实验组 1(生理盐水 )、实验组 2 (碘化油 )、实验组 3(As2 O3 +碘化油 )经肝动脉注射治疗肝移植瘤一次 ,实验组 4 (As2 O3 )经肝动脉插管灌注药物 (连续 7天 )。观察肿瘤平均瘤重、平均瘤重抑制率、肿瘤细胞形态学变化 ,检测bcl 2 /bax基因表达和VEGF表达。结果　实验组平均瘤重分别为 16 .0 8± 2 .6 9g、3.4 4± 0 .84 g、1.5 2± 0 .2 6 g和4 .87± 0 .73g ,平均瘤重抑制率分别为 78.6 1%、90 .5 5 %和 6 9.71% ,实验组 3移植瘤瘤重低于其他实验组 (P <0 .0 5 )。实验组 3、4移植瘤组织bax基因表达显著上调、bcl 2基因表达显著下调 (P <0 .0 5 )、VEGF表达下调 (P <0 .0 5 ) ;电镜观察实验组 3、4均有典型的瘤细胞凋亡改变。结论　As2 O3 经肝动脉栓塞化疗对兔Vx 2肝移植瘤有显著抑瘤作用 ,碘化油能增强As2 O3 对肝移植瘤的抑制作用 ,两药具有协同作用。     

组织块悬液注射法制作兔VX2乳腺癌模型

目的：建立稳定的兔VX2乳腺癌模型，选择最佳的模型制作方法。方法：30只兔随机分为3组，每组10只，分别采用乳腺内瘤细胞液注射法、组织块悬液注射法、组织块种植法制作兔VX2乳腺癌模型，比较各组的种植成活率、肿瘤生长率、荷瘤兔存活时间，并观察肿瘤的生物学形态。结果：组织块悬液注射法操作简便，成瘤率高，肿瘤增殖旺盛，生物学特征符合乳腺鳞状细胞癌。结论：成功建立了兔VX2乳腺癌模型，组织块悬液注射法是最佳模型制作方法。     

白头翁醇提物抗荷瘤鼠肿瘤血管生成作用的实验研究

目的:研究白头翁醇提物对昆明小鼠H22肝癌细胞皮下移植瘤的抗血管生成作用。方法:建立鼠H22肝癌细胞小鼠皮下移植瘤模型,给予不同剂量白头翁醇提物灌胃治疗,连续用药10天后,观察用药后肿瘤的大小,计算瘤重抑制率并测定肿瘤微血管密度(MVD)。结果:20g/kg白头翁醇提物对小鼠H22细胞皮E移植瘤作用后瘤重显著低于对照组(P=0.01)。10g/kg、20g/kg白头翁醇提物对小鼠H22细胞皮下移植瘤治疗后的微血管计数均显著低于对照组(P值分别为0.037、0.01)。结论:白头翁醇提物对荷瘤鼠H22肝癌移植瘤具有抗肿瘤作用,其作用机理可能与抗肿瘤血管生成作用有关。     

Immune elimination of host fibroblasts for the cultivation of human tumors transplanted into nude mice.

We report here a useful method for the isolation and cultivation of human tumor cells in vitro from human tumors grown in nude mice. A rabbit was immunized with spleen cells obtained from adult nude mice. The rabbit antiserum in the presence of complement effectively killed cultured cells derived from various mouse tissues, but it was not cytotoxic to cultured cells from human tissues including tumors. When mixed cultures consisting of human tumor cells and nude mouse fibroblasts were treated with the antiserum and complement, the nude mouse fibroblasts were completely removed from the cultures, and the human tumor cells could be propagated without noticeable changes in morphological features. Primary cultures of heterotransplanted human tumors grown in nude mice were also successfully treated, resulting in the ultimate elimination of fibroblastic cells derived from the stroma of the tumor. The functional properties of the tumor cells (production of human chorionic gonadotropin by choriocarcinoma cells and production of carcinoembryonic antigens by pancreas carcinoma cells) were also maintained after the antiserum treatment.     

内皮抑素联合微波消融治疗裸鼠肾癌移植瘤研究

目的研究内皮抑素联合微波消融对裸鼠肾癌移植瘤的抑制作用。方法将人源性肾癌细胞接种到48只小鼠的背部皮下,肿瘤直径约1.5 cm时随机分成4组,对照组：隔日尾静脉生理盐水;联合治疗组：隔日尾静脉推注内皮抑素＋皮下肿瘤微波消融;微波消融组：皮下肿瘤微波消融;内皮抑素组：隔日尾静脉推注内皮抑素。首先对联合治疗组及微波消融组局部皮下肾移植瘤进行微波消融治疗,其后经尾静脉注射药物。14天后每组处死6只小鼠,比较各组瘤重,计算抑瘤率,检测血浆VEGF浓度,剩余6只小鼠观察其存活期。结果联合治疗组小鼠肿瘤体积显著小于其他各组（P〈0.0 5）,其血浆VEGF水平较其他各组显著减低（P〈0.0 5）,存活时间较其他各组显著延长（P〈0.05）。结论内皮抑素联合微波消融抗肿瘤作用优于单用内皮抑素或微波消融治疗,并使小鼠存活期延长。     

Renal cell carcinoma from a transplanted allograft: two case reports and a review of the literature

We report 2 cases of renal cell carcinoma (RCC) in which the tumor arose from a transplanted allograft. The first case is a 52-year-old man with a failed cadaveric renal transplantation found to have metastatic RCC. The tumor was proven to be from the allograft, as fluorescence in situ hybridization analysis of biopsy material showed a female karyotype, consistent with his female donor. The second patient is a 45-year-old man who had undergone cadaveric renal transplantation in 1985 for chronic glomerulonephritis and, after 22 years, presented with renal failure. Biopsy and subsequent allograft nephrectomy revealed innumerable microscopic foci of RCC. There are only a few reported cases of RCC arising in kidney allografts and even fewer with reports of metastatic disease from the allograft. Treatments in patients with disease confined to the kidney have included partial nephrectomy and total nephrectomy. A literature search did not find any reports of treatment of metastatic RCC that arose from a renal allograft.     

Molecular imaging of angiogenesis in nascent Vx-2 rabbit tumors using a novel alpha(nu)beta3-targeted nanoparticle and 1.5 tesla magnetic resonance imaging

Early noninvasive detection and characterization of solid tumors and their supporting neovasculature is a fundamental prerequisite for effective therapeutic intervention, particularly antiangiogenic treatment regimens. Emerging molecular imaging techniques now allow recognition of early biochemical, physiological, and anatomical changes before manifestation of gross pathological changes. Although new tumor, vascular, extracellular matrix, and lymphatic biomarkers continue to be discovered, the alpha(nu)beta(3)-integrin remains an attractive biochemical epitope that is highly expressed on activated neovascular endothelial cells and essentially absent on mature quiescent cells. In this study, we report the first in vivo use of a magnetic resonance (MR) molecular imaging nanoparticle to sensitively detect and spatially characterize neovascularity induced by implantation of the rabbit Vx-2 tumor using a common clinical field strength (1.5T). New Zealand White rabbits (2 kg) 12 days after implantation of fresh Vx-2 tumors (2 x 2 x 2 mm(3)) were randomized into one of three treatment groups: (a) alpha(nu)beta(3)-targeted, paramagnetic formulation; (b) nontargeted, paramagnetic formulation; and (c) alpha(nu)beta(3)-targeted nonparamagnetic nanoparticles followed by (2 h) the alpha(nu)beta(3)-targeted, paramagnetic formulation to competitively block magnetic resonance imaging (MRI) signal enhancement. After i.v. systemic injection (0.5 ml of nanoparticles/kg), dynamic T(1)-weighted MRI was used to spatially and temporally determine nanoparticle deposition in the tumor and adjacent tissues, including skeletal muscle. At 2-h postinjection, alpha(nu)beta(3)-targeted paramagnetic nanoparticles increased MRI signal by 126% in asymmetrically distributed regions primarily in the periphery of the tumor. Similar increases in MR contrast were also observed within the walls of some vessels proximate to the tumor. Despite their relatively large size, nanoparticles penetrated into the leaky tumor neovasculature but did not appreciably migrate into the interstitium, leading to a 56% increase in MR signal at 2 h. Pretargeting of the alpha(nu)beta(3)-integrin with nonparamagnetic nanoparticles competitively blocked the specific binding of alpha(nu)beta(3)-targeted paramagnetic nanoparticles, decreasing the MR signal enhancement (50%) to a level attributable to local extravasation. The MR signal of adjacent hindlimb muscle or contralateral control tissues was unchanged by either the alpha(nu)beta(3)-targeted or control paramagnetic agents. Immunohistochemistry of alpha(nu)beta(3)-integrin corroborated the extent and asymmetric distribution of neovascularity observed by MRI. These studies demonstrate the potential of this targeted molecular imaging agent to detect and characterize (both biochemically and morphologically) early angiogenesis induced by minute solid tumors with a clinical 1.5 Tesla MRI scanner, facilitating the localization of nascent cancers or metastases, as well as providing tools to phenotypically categorize and segment patient populations for therapy and to longitudinally follow the effectiveness of antitumor treatment regimens.