人前列腺癌裸鼠皮下移植瘤模型的建立

目的 建立人前列腺癌裸鼠皮下移植瘤模型.方法 采用人前列腺癌细胞株PC-3细胞接种于裸鼠的颈背部皮下,计算成瘤潜伏期、成瘤百分率,观察移植瘤大体生长情况,绘制生长曲线,并对移植瘤进行病理鉴定.结果 采用人前列腺癌细胞株PC-3细胞皮下接种方式建立移植瘤的平均成瘤潜伏期为24天,成瘤百分率为100％,瘤体积倍增时间为10天左右,移植瘤的形态和功能特性与原发肿瘤基本相似.结论 本动物模型的建立可为前列腺癌的放射免疫显像以及放射免疫治疗提供一个有价值的实验平台.     

人绒毛膜癌裸鼠原位移植模型的建立

目的建立人绒毛膜癌裸鼠原位移植瘤模型。方法培养人绒毛膜癌细胞系JAR,制备JAR单细胞悬液,给5只8周龄BALB/c裸鼠经皮下注射建立皮下移植瘤模型。待裸鼠皮下成瘤后,无菌条件下取瘤组织并切成1 mm^3组织块,通过手术方式植入10只10周龄BALB/c裸鼠子宫腔内,裸鼠濒死状时4%水合氯醛(10 g/kg)腹腔注射麻醉处死,观察子宫成瘤及腹腔转移情况。解剖取子宫原位移植瘤、腹腔内转移瘤、腹腔淋巴结及其他脏器组织标本,通过组织病理学检查进行鉴定。结果10只BALB/c裸鼠中共有7只裸鼠子宫内可见移植瘤肿块形成,其中2只可同时观察到子宫移植瘤和腹膜转移瘤。在病理学形态和结构上,皮下移植瘤模型、原位移植模型和腹膜转移瘤的瘤细胞与人绒毛膜癌细胞系JAR一致。结论成功建立人绒毛膜癌JAR细胞的BALB/c裸鼠原位移植瘤模型。     

裸鼠皮下人肝癌移植瘤模型的建立

目的建立裸鼠皮下肝癌移植瘤模型。方法BLBA/c裸鼠颈背部皮下注射人肝癌SMMC-7721细胞悬液5×106个.0.2m l-1.只-1,观察肿瘤生长情况,45d处死。瘤组织作病理及电镜检查;取裸鼠周围血作甲胎蛋白(AFP)的检测;用免疫组化法检测肿瘤微血管密度(MVD)。结果该模型具有类似人原发性肝癌的形态学特征。结论成功建立了人肝癌裸鼠皮下移植瘤模型。     

采用组织块移植法快速建立裸鼠原位骨肉瘤模型

背景：骨肉瘤细胞起源于间叶组织这一特点使得骨肉瘤动物模型的构建存在较多困难,进展缓慢的问题。 目的：采用组织块移植法建立裸鼠原位骨肉瘤模型,并观察和评价其生物学特性。 方法：选用SPF 级 6 周龄裸鼠15只,将制备好的MG-63 骨肉瘤细胞悬液注射到裸鼠右前肢腋下,成瘤后连续传 3 代,待肿瘤生长稳定后将瘤块组织移植到裸鼠下肢胫骨髓腔内,评价其生物学特性,采用X射线观察裸鼠成瘤率和成瘤特点。并以正常裸鼠5只做对照。 结果与结论：15只裸鼠造模过程顺利,造模后通过观察1只成瘤失败,成瘤率为93%;造模后观察发现采用组织块移植法三四周可见裸鼠下肢局部肿瘤状组织形成,四五周后X射线观察可见裸鼠胫骨中上段出现不明显的溶骨以及瘤样骨形成;骨肉瘤模型组碱性磷酸酶的表达水平明显高于正常对照组(P < 0.01)。数据表明采用组织块移植法进行裸鼠原位骨肉瘤造模,其造模方法简单,成瘤率高,瘤体生长速度快,对骨皮质及周围软组织的破坏力较强,接近临床骨肉瘤患者的实际情况。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程      

稳定表达hTERT/Luc的小鼠黑色毒瘤肺转移模型的建立

目的 建立稳定共表达荧光素酶基因和人端粒酶逆转录酶(hTERT)的小鼠黑色素瘤B16细胞系,并通过尾静脉注射的方式建立小鼠肿瘤肺转移模型.方法 利用DNA重组技术将hTERT基因和荧光素酶基因Luc定向插入到真核表达载体,构建真核表达质粒pIRES-neo-hTERT和pIRES-hyg3-Luc,利用阳离子脂质体LipofectamineTM 2000共转染小鼠黑色素瘤B16细胞,经G418及潮霉素B加压筛选出稳定转染的细胞株.应用Western blot法及免疫荧光法检测hTERT和Luc基因在B16细胞中的表达;将稳定共表达hTERT和Luc的B16-hTERT/Luc细胞株通过尾静脉注射的方式接种雄性C57BL/6小鼠建立肿瘤肺转移模型,并通过活体成像技术检测小鼠肺部肿瘤的生长.结果 建立了稳定共表达hTERT和Luc的小鼠黑色素瘤B16单克隆细胞株B16-hTERT/Luc,经检测hTERT基因和荧光素酶基因Luc在单克隆细胞株中的表达分别为84％和98％.通过尾静脉注射的方式成功建立了小鼠肿瘤肺转移模型,应用活体成像技术能方便地检测到B16-hTERT/Luc肿瘤在小鼠体内的生长情况.结论 成功建立了可用于活体成像技术检测的稳定表达hTERT的小鼠黑色素瘤肺转移模型.     

两株肺癌细胞系体外培养细胞和裸鼠体内移植瘤细胞增殖状态的比较研究

目的：比较两株肺癌细胞系（经实验证明为高、低侵袭系）体外培养细胞和裸鼠体内移植瘤细胞的增殖状态；方法：采用激光流式细胞仪，测量单个完整细胞群体的ＤＮＡ含量，分析其各细胞周期的比率；结果：体外培养的高增殖系细胞，移植入裸鼠体内后其Ｇ２／Ｍ期细胞比率反而降低，体外培养的低增殖系细胞，移植入裸鼠体内后其Ｇ２／Ｍ细胞比率反而升高，两株肺癌细胞系的增殖指数间无显著差异；结论：两株肺癌细胞系在体内、外增殖状况的差异性，说明肿瘤细胞在体外增殖速度的快慢不能准确反映其在体内的增殖状态，必须结合肿瘤细胞的其它生物学特性，才能比较准确地分析和判断肿瘤细胞的恶性程度。     

端粒酶核酶抑制裸鼠移植瘤生长的实验研究

目的　探讨端粒酶核酶对裸鼠移植瘤生长的影响。方法　构建了人子宫颈癌细胞株HeLa 裸鼠移植瘤,将不同剂量端粒酶核酶真核表达质粒pXJ-neo-teloRZ用脂质体包裹后进行瘤体内多点注射,对照组注射生理盐水或空质粒载体。连续注射14 天,测量肿瘤体积和重量,检测瘤组织端粒酶活性,并作病理切片检查。结果　端粒酶核酶使肿瘤组织端粒酶活性下降,肿瘤组织凋亡增加,明显地抑制了裸鼠移植瘤生长。结论　该端粒酶核酶可望成为有效的端粒酶抑制剂,在肿瘤基因治疗中发挥作用。     

载异种移植瘤的裸鼠Ig分泌细胞的检测

裸鼠对子异种移植肿瘤不能产生有效的免疫排斥反应。为了观察裸鼠对异种移植肿瘤是否能产生其它类别的免疫应答,本文用葡萄球菌蛋白A空斑形成细胞检测法对6只健康的BALB/c裸鼠和9只载有人类移植瘤的裸鼠的脾和骨髓作了免疫球蛋白分泌细胞(IgSC)的检测。结果表明,载瘤裸鼠脾和骨髓中IgSC的数量分别高于健康裸鼠相应部位的IgSC数量。载瘤与未载瘤两组裸鼠骨髓中每10~5个有核细胞的IgSC数分别为72±21和7.8±2.2,两数的差别有显著性(t=6.48,P<0.001)。载瘤裸鼠骨髓IgSC的增加显然是由于肿瘤细胞的刺激引起的。     

人直肠粘液腺癌裸鼠移植瘤模型的建立及其生物学特性

A model of transplantable human mucoid adenocarcinoma of rectum in BALB/C nu/nu nude mice (TNB 92), was established and 18 sub-transplantations were performed. The success rate of transplantation was 98% and the average tumor bearing life time of mice was 112 days. Histology and ultrastructures showed that the transplantable tumor retained the original structures of the human tumor. Chromosomal analysis of the tumor cells exhibited the same features of the human carcinoma and it also retained the function of secreting CEA. Frozen tissue of the tumor in liquid nitrogen after rehabilitation could be successfully retransplanted into nude mice again. It seems to be a useful model for further study of human rectal adenocarcinoma.     

NK4基因转染对裸鼠人淋巴瘤移植瘤的抑制作用

目的 探讨NK4基因转染对裸鼠人淋巴瘤移植瘤的抑制作用及其机制.方法 采用NK4基因重组质粒pVITRO2-NK4转染的Raji细胞建立裸鼠皮下人淋巴瘤移植瘤模型,动态监测裸鼠体重和肿瘤大小.8周后获取瘤组织,分别采用免疫组化和脱氧核糖核酸末端转移酶介导的缺口末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)检测移植瘤组织的细胞凋亡和微血管密度(microvessel density,MVD),并进行相关分析.结果 NK4基因转染组裸鼠移植瘤体积明显小于对照组(P<0.01),而对照组间差异无显著性(P>0.05);各组间裸鼠体重降低,差异无显著性(P<0.01).NK4基因转染组的淋巴瘤细胞AI值达237±10.94,而质粒pVITRO2转染组和未转染Raji组的AI值分别为79.7±30.8和81.7±22.2,NK4基因转染组明显高于对照组(P<0.001),而对照组间差异无显著性(P>0.05);NK4基因转染组MVD为4.7±1.52,而质粒pVITRO2转染组和未转染Raji组MVD分别为12.0±1.00和10.66±1.53,NK4基因转染组明显低于对照组(P<0.001),而对照组间差异无显著性(P>0.05).结论 NK4基因转染可明显抑制裸鼠人淋巴瘤移植瘤的生长,其可能通过抑制肿瘤血管新生和促肿瘤细胞凋亡而发挥其效应.     

食管癌耐药皮下移植瘤裸鼠模型建立

 [摘要] 目的 建立食管癌耐药裸鼠模型及探讨食管癌耐药机制。方法 4周龄的BALB/c nu/nu裸小鼠36只随机分组分为6组,每组6只,左前肢肩胛下皮下分别接种食管癌细胞Eca109及食管癌耐药细胞Eca109/ABCG2,建立裸鼠皮下移植瘤模型。成瘤后腹腔注射阿霉素（Adriamycin, ADM）,1、4mg/kg,1次/3d 共注射7次,空白对照组使用生理盐水(normal saline, NS)代替ADM。RT-PCR方法检测移植瘤细胞中三磷酸腺苷结合转运蛋白G2（ATP-binding cassette transporter G2,ABCG2）mRNA 表达情况,流式细胞术(Flow cytometry, FCM)检测移植瘤细胞中ABCG2蛋白、凋亡及细胞中ADM含量。结果 成功建立裸鼠食管癌耐药细胞移植瘤模型,皮下接种细胞一周后成瘤,成瘤率100%。实验结束后,注射ADM药物的裸鼠,接种Eca109/ABCG2细胞的皮下移植瘤体积、重量和ABCG2 mRNA、蛋白表达量显著高于接种Eca109细胞移植瘤（P<0.05）,但细胞凋亡率及细胞内ADM含量显著降低（P<0.05）。结论 接种Eca109/ABCG2细胞的裸鼠皮下移植瘤模型是较好的食管癌耐药动物模型,具有ABCG2耐药表型,为研究ABCG2与食管癌耐药关系提供了较理想的动物模型。     

Establishment and studies of two human pancreatic carcinoma in vivo lines by transplanting the tumor into nude mice

Two human pancreatic adenocarcinoma in vivo lines (PCT-1, PCT-2) were established in athymic nude mice and maintained for 27 passages (PCT-1) and 7 passages (PCT-2) respectively in 29 (PCT-1) and 10.5 months (PCT-2). Tumours obtained from metastatic lymph node (PCT-1) and metastatic nodule in the omentum (PCT-2) were the initial sources for implantation. All xenografts in the two tumour lines were studied by using histological, ultrastructural, immunohistochemical and cytogenetical methods. PCT-1 maintained the poorly differentiated adenocarcinoma features with little mucin production, and CEA was weakly positive with metastasis to lymph nodes in this tumour line. PCT-2 was a moderately differentiated adenocarcinoma line with abundant mucin secretion, CEA was strongly positive. The poorly differentiated tumours have a faster growth rate in comparison with those of the moderately and well differentiated ones.     

Combined MR,fluorescence and histology imaging strategy in a human breast tumor xenograft model

Applications of molecular imaging in cancer and other diseases frequently require the combination of in vivo imaging modalities, such as MR and optical imaging, with ex vivo optical, fluorescence, histology and immunohistochemical imaging to investigate and relate molecular and biological processes to imaging parameters within the same region of interest. We have developed a multimodal image reconstruction and fusion framework that accurately combines in vivo MRI and MRSI, ex vivo brightfield and fluorescence microscopic imaging and ex vivo histology imaging. Ex vivo brightfield microscopic imaging was used as an intermediate modality to facilitate the ultimate link between ex vivo histology and in vivo MRI/MRSI. Tissue sectioning necessary for optical and histology imaging required the generation of a three‐dimensional reconstruction module for two‐dimensional ex vivo optical and histology imaging data. We developed an external fiducial marker‐based three‐dimensional reconstruction method, which was able to fuse optical brightfield and fluorescence with histology imaging data. The registration of the three‐dimensional tumor shape was pursued to combine in vivo MRI/MRSI and ex vivo optical brightfield and fluorescence imaging data. This registration strategy was applied to in vivo MRI/MRSI, ex vivo optical brightfield/fluorescence and histology imaging datasets obtained from human breast tumor models. Three‐dimensional human breast tumor datasets were successfully reconstructed and fused with this platform. Copyright © 2012 John Wiley & Sons, Ltd.     

大鼠胰岛素瘤实验模型的建立及其应用

目的 通过移植大鼠胰岛素瘤INS-1细胞至糖尿病小鼠肾包膜下使之形成胰岛素瘤,并诱导其发生凋亡,建立可用于研究胰岛β细胞凋亡机制的动物模型.方法 将5×106 INS-1细胞接种于链脲霉素(STZ)造模的糖尿病小鼠左肾包膜下,监测动物空腹血糖和血清胰岛素水平的变化,当血糖趋近正常后摘取动物左侧肾脏并检测其血糖和血清胰岛素水平的变化;固定包埋摘取的肾脏,进行HE染色以及胰岛素的免疫组化染色,确定胰岛素瘤模型的建立.在胰岛素瘤动物模型中,腹腔给予毒胡萝卜素(TG)或软脂酸钠(PA),监测给药后动物空腹血糖的变化,当血糖浓度出现逆转时,摘取动物左侧肾脏,通过TUNEL原位染色法检测移植瘤细胞的凋亡.结果 将INS-1细胞移植到糖尿病小鼠肾包膜下后,从第9天开始,动物空腹血糖进行性降低,血清胰岛素水平逐渐升高,当动物血糖接近至正常时,摘取动物左肾导致动物血糖显著升高,在摘取的左肾可见明显的移植瘤,免疫组织化学染色显示移植瘤细胞为胰岛素阳性.在胰岛素瘤动物模型给予TG或PA刺激后,动物空腹血糖出现逆转,显著升高,血清中胰岛素含量明显降低,摘取动物左侧肾脏后,TUNEL原位染色发现移植瘤内有明显的细胞凋亡.结论 大鼠胰岛素瘤INS-1细胞肾包膜下移植可以建立胰岛素瘤动物模型,应用此动物模型可以在体内研究胰岛β细胞凋亡的机制.     

RNA干扰下调Stat5对人肝癌裸鼠移植瘤的抑制及诱导肿瘤细胞凋亡的研究

目的 探讨Stat5-shRNA对裸鼠体内人肝癌SMMC7721移植瘤的抑制及其诱导肿瘤细胞凋亡的作用.方法 将SMMC7721细胞悬液接种于裸鼠背部皮下,15d后,待在接种部位出现肿瘤结节、质地较硬等指标认定为成瘤.将15只成瘤裸鼠完全随机分为:空白对照组、HK质粒对照组、Stat5-shRNA组共3组,每组各5只.空白对照组瘤内注射生理盐水,HK质粒对照组瘤内注射Pgenesil1-HK,Stat5-shRNA组瘤内注射Pgenesil-1 -Stat5A1,各组注射剂量及次数均为50μl/只,每2天1次,共10次.第35天,处死各组全部动物,剥瘤称重,计算肿瘤大小及抑瘤率,流式细胞术检测肿瘤细胞凋亡情况.结果 与空白对照组和HK质粒对照组比较,Stat5-shRNA组裸鼠人肝癌移植瘤的生长受到明显抑制,肿瘤细胞凋亡率明显上升[(21.35±3.69)％比(3.56±1.12)％,(3.81±3.05)％,P＜0.05].结论 Stat5-shRNA可有效抑制裸鼠体内人肝癌移植瘤的生长,并能够诱导肿瘤细胞凋亡.Stat5-shRNA在人肝癌的基因治疗中具有潜在的应用价值.     

KLF17对裸鼠移植瘤生长的影响及其在体调控靶基因的筛选和功能分析

目的:研究Krüppel样因子17(Krüppel-like factor 17,KLF17)在裸鼠移植瘤中的作用并分析KLF17在体调控的靶基因及其功能和参与的信号通路。方法:慢病毒转染技术稳定上调人肺腺癌A549细胞及下调人肺腺癌H322细胞中KLF17的表达。11只BLAB/c nu/nu裸鼠分为上调组5只和下调组6只,分别通过左、右侧躯体皮下注射KLF17上调和下调的相应细胞及其对照细胞,观察KLF17对裸鼠移植瘤生长的影响;Real-time PCR及免疫组织化学染色分析裸鼠移植瘤组织中KLF17 mRNA及蛋白的表达,转录组测序技术分析上调KLF17表达后裸鼠移植瘤中差异表达的基因,通过The Cancer Genome Atlas(TCGA)数据库分析KLF17调控的差异基因的功能,Gene Ontology和KEGG PATHWAY富集分析差异基因的功能和可能参与的信号通路。结果 :上调A549细胞中KLF17的表达后,其裸鼠移植瘤生长速率显著低于空载体对照组(P0.05),下调H322细胞中KLF17表达后,其裸鼠移植瘤生长速率及移植瘤重量均显著高于空载体对照组(P0.01,P0.05)。在裸鼠移植瘤组织中,过表达组KLF17 mRNA及蛋白显著高于对照空载体组。转录组测序结果显示KLF17可能调控的基因有ras基因同源家族成员V(ras homolog family member V,RHOV)和冠蛋白1C(coronin 1C,CORO1C)等。TCGA数据库中肺腺癌患者10年累计生存时间在RHOV和CORO1C mRNA不同表达组明显不同,高表达RHOV及CORO1C的肺腺癌患者生存时间显著低于其低表达组。Gene Ontology和KEGG PATHWAY富集分析提示KLF17调控的靶基因(差异基因)可能与肿瘤细胞的刺激应答、生长及黏附有关,并且参与了细胞趋化性、黏附及细胞外基质受体相关的信号通路。结论:KLF17抑制裸鼠移植瘤的生长,并下调RHOV和CORO1C等癌基因,参与调控肿瘤黏附及生长相关信号通路。     

Enhancing magnetic resonance imaging tumor detection with fluorescence intensity and lifetime imaging

Early detection is important for many solid cancers but the images provided by ultrasound, magnetic resonance imaging (MRI), and computed tomography applied alone or together, are often not sufficient for decisive early screening ∕ diagnosis. We demonstrate that MRI augmented with fluorescence intensity (FI) substantially improves detection. Early stage murine pancreatic tumors that could not be identified by blinded, skilled observers using MRI alone, were easily identified with MRI along with FI images acquired with photomultiplier tube detection and offset laser scanning. Moreover, we show that fluorescence lifetime (FLT) imaging enables positive identification of the labeling fluorophore and discriminates it from surrounding tissue autofluorescence. Our data suggest combined-modality imaging with MRI, FI, and FLT can be used to screen and diagnose early tumors.     

Establishment of fluorescent lung carcinoma metastasis model and its real-time microscopic detection in SCID mice

Lung cancer is the most prevalent malignant tumor in the world. Metastasis of the disease causes death in lung cancer patients. Recent study has shown that multiple cascades of gene defects occur in lung cancer. In this report, we established a novel H1299/EGFP tumor model to determine whether H1299 transfected with the enhanced green fluorescent protein (EGFP) gene in vitro and xenotransplanted into SCID mouse lung would permit the detection of lung cancer micro-metastasis in vivo. We demonstrated that EGFP-transduced H1299 cells maintained stable high-level EGFP expressions during their growth in vivo. EGFP fluorescence clearly demarcated the primary seeding place and readily allowed for the visualization of distant micrometastasis and local invasion at the single-cell level. Small metastatic and locally invasive foci, including those immediately adjacent to the tumor's leading invasive edge, were almost undetectable by routine hematoxylin and eosin staining and immunohistochemistry. The GFP tagged lung cancer model is superior for the detection and study of physiologically relevant patterns of lung cancer invasion and metastasis in vivo. This revised version was published online in July 2006 with corrections to the Cover Date.