We thank Dr. Roger Erickson for his support and assistance with the preparation of the manuscript. This work was supported by the National Cancer Institute at the National Institutes of Health through grant numbers RO1CA160079, RO1CA138642, UO1CA184966 and VA funded program project number 1P1 BX001604.

涉及动物的所有程序协议必须经过审核并通过机构动物护理和使用委员会（IACUC）的批准。按照照顾和使用实验动物的正式批准程序。帧内前列腺注射需要开腹部手术和动物应与在适当的手术无菌技术在整个过程期间使用的指定手术室保持在无病原体的环境中。 1.细胞植入的制备注意:基于研究的需要，可使用任何前列腺癌细胞系。细胞系是根据供应商的说明进行培养。 <ol><li>对于稳定表达萤火虫萤光素酶基因的PC3M吕克-C 6细胞系，在最低必需培养基（MEM）中培养的细胞在补充有10％胎牛血清（FBS），1×非必需氨基酸，1×腐草霉素D1和1mM的丙酮酸钠。维持细胞在培养箱瓦特第i个95％空气和5％ 的 CO 2在37℃的湿润气氛中。 PC3M - 吕克·-C6细胞从加州大学旧金山分校的核心设备采购。 </li><li>收获细胞胰蛋白酶消化。用磷酸盐缓冲盐水（PBS）洗涤培养板一次。用95％空气和5％CO 2的潮湿气氛中在37℃下加2ml的0.05％胰蛋白酶至10cm皿并孵育3-5分钟的培养箱直到细胞分离。 <ol><li>为了避免结块，不通过敲击或振动的菜，而等待的细胞分离搅动细胞。收集细胞在5ml完全培养基，并降速在200×g下5分钟。用PBS洗涤细胞沉淀，除去胰蛋白酶。 </li></ol></li><li>通过台盼蓝排除法一一列举活细胞。混合10微升在PBS中的细胞悬浮液与0.4％的10微升（重量/体积）台盼蓝溶液。加载混合物成血球或计数室滑动件和在显微镜下立即计数细胞或阅读在细胞凑腔室滑动NTER。 <ol><li>制备含有2.5×10 5个细胞于10微升培养基的细胞悬浮液。混合10微升基底膜样胞外基质提取物中的细胞悬浮液，然后将细胞在冰上。注射到小鼠中之前;（库存30毫克/毫升浓度200μl的1）添加荧光素的细胞悬浮液。 注意:这允许动物的即时生物成像检查不同的实验组之间的细胞注射的一致性。注入细胞尽快，优选在30分钟胰蛋白酶消化后自细胞活力脱离后迅速下降。 </li></ol></li></ol> 2.手术区的制备<ol><li>在促进无菌整洁，消毒面积进行手术。 </li><li>消毒术前漂白粉溶液计数器/实验室工作台。 注意:使用酒精是泄气因生效（15分钟）所需的接触时间长。 </li><li>使用无菌单，清洁吸收垫或毛巾，每个手术后会更换这些材料。消毒，使用前所有仪器。优选的方法是蒸汽高压釜中，玻璃珠灭菌，环氧乙烷气体，或过氧化氢蒸汽消毒。 </li><li>使用清洁无菌解剖显微镜进行外科手术或有经验的研究人员可以不用显微镜执行它。 </li></ol> 3.肿瘤细胞移植<ol><li>男性使用6-8周龄免疫缺陷的Balb / c或NOD / SCID小鼠。 注:它是对较小的动物，以及较大的动物操作更加困难往往具有肿瘤生长和转移的较慢动力学。 </li><li>根据动物设施的指令注入手术前的止痛药。例如，可使用的剂量为0.1毫克/公斤体重腹膜内丁丙诺啡。 </li><li>通过在氧和WA将它们放置在一个异氟烷室用1-3％异氟烷麻醉动物直到动物完全麻醉。确保有在这一点上的肌肉张力的无趾反射。使用适当的兽医眼药膏，防止润滑油全身麻醉期间，由于干眼症失明。 注意:由研究者的优选方法例如麻醉动物，对于戊巴比妥钠，0.05毫克每克体重施用腹膜内或氯胺酮/甲苯噻嗪溶液（浓度:17.16毫克/毫升），剂量为65毫克/千克体重经皮下使用。异氟醚麻醉是一个首选方法。该眼药膏应无摩擦角膜被轻轻涂抹。 </li><li>从异氟烷室取出动物并放入在氧气1-2％异氟醚的连续流动的鼻锥装置，以确保动物是全身麻醉下实施，然后再继续。 <ol><li>通过剃须删除小鼠毛发或开始操作之前，使用脱毛膏。 注:这将是最好放置在无菌加热垫鼠标手术过程中。 </li></ol></li><li>放置鼠标在仰卧位置。清洗，用10％的下腹部W W，随后用70％乙醇拭子/聚维酮碘溶液。 </li><li>有一对细镊子，抬起的皮肤区域的包皮接头2毫米的位置，约1-2厘米的阴茎鞘的上方，并且肋笼的底部低于约2-3厘米。 </li><li>使一个中线切口1cm的长度，首先通过用解剖刀的皮肤，然后通过肌肉层用剪刀（ 图1）。 </li><li>定位在体腔内的膀胱。它是一种黄色的，浅棕色球形器官，直接位于切口（ 图1）下。 </li><li>有一对细钳子把持的膀胱和向下抬起向上再出朝向阴茎鞘体腔。这将会使两个精囊这是一双白色囊状器官和明显不同。 </li><li>随着棉签每手，外部化精囊，一个接一个，拉出来的体腔，躺在它们面朝下在中间膀胱（ 图2）腹部的外表面。 </li><li>使用棉签轻轻在膀胱颈附近插入点向后倾斜的精囊，朝向阴茎鞘，使两背侧前列腺叶都清晰可见。用湿棉签，以避免组织损伤（ 图3）。 </li><li>在装入注射器搅拌前用微量细胞悬液。 </li><li>而用棉签将精囊中的位置，将注射器针头插入显微镜下背侧前列腺叶（ 图4）。缓缓注入20微升细胞悬浮液，直到泡的形成被识别。一个鼓鼓的大泡表明，注射是正确的。 </li><li>同时轻轻缩回针头，按注射部位上棉签并保持几秒钟，以防止泄漏。 </li><li>小心地提起用棉签精囊和由一个后跟膀胱插回到体腔之一。避免"捻转&#39;内脏，而执行此过程。 </li><li>放置器官回进入体腔后，在用可吸收4-0铬肠线缝合接着皮肤缝合以非吸收性4-0尼龙手术缝线断续图案的第一缝合肌肉层。皮肤也能够一起拉出并与手术夹子闭合以完全关闭切口。 注:小鼠有划伤和在它们的伤口，这可能会导致重新开口的伤口，因此，使用组织胶的咬的习惯，还建议连同缝线。图像动物立即以确保有各实验组之间的平等生物发光的强度。 </li><li>返回动物清洁笼子，并保持他们变暖灯或下热ING垫。监视动物不断，直到它们完全从麻醉中恢复并保持胸骨斜卧。 </li></ol> 4.动物监测<ol><li>定期监测这些动物直到试验结束时，根据机构的协议。如果使用的手术金属夹，一到两周后拆下。实验的持续时间取决于具体的研究需要。 注:此实验21天进行，探讨在小鼠前列腺成功建立植入癌细胞。 </li><li>管理基于所述动物设施的指示的止痛药。例如，在剂量为0.1毫克/公斤体重腹膜内丁丙诺啡可以在与第二剂量的步骤的时间后6小时和附加剂量根据需要使用的每8-12小时。 </li><li>监测动物体重，食品消费，皮肤颜色和质地，活动及排尿和排便次数。动物实施安乐死IMMEDiately如果有体重大于15％的显著损失。 <ol><li>安乐死，在流速从加压罐递送的 CO 2进入的未挤笼来置换腔室或笼体积/分钟的10-30％，从而使无意识和完整narcotization发生的 CO 2慢慢进入腔室之前死亡。 </li><li>维持二氧化碳流为呼吸停止后至少一分钟，离开动物在腔室足够长的时间，使得前执行物理方法时发生的死亡。 </li><li>执行斩首，颈椎脱位或任何其他IACUC化学安乐死动物批准后的物理方法。 注:显着降低体重常提示病情昏昏欲睡。荷瘤动物应该在良好卫生除了肿瘤的存在，直到实验结束。安乐死应该是关于安乐死的AVMA指引一致，且不能清单当然d。在批准IACUC协议。 </li></ol></li></ol>动物5.非侵入性生物成像<ol><li>监测动物每周使用非侵入性成像技术来跟踪癌细胞，肿瘤的生长和任何远处转移的定植。 注:可以基于特定的研究中使用成像方式如GFP成像，萤光素酶的成像，X射线或3D显微计算机断层摄影术（UCT） 等需要12-15。 </li></ol>

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作者宣称，他们没有竞争的经济利益。

这份手稿描述了建立人体原位前列腺癌异种移植小鼠模型的详细过程。该模型是由人前列腺癌细胞系PC3M吕克-C 6的直接植入建立到免疫缺陷小鼠的背侧前列腺叶。肿瘤被允许开发在实验的过程中。在实验过程中的肿瘤生长每周监测由非侵入性生物发光成像系统。 在建立异种移植肿瘤模型中最重要的因素是整个肿瘤细胞的植入实现的一致性。以获得统计学显著的结果，各试验组...

前列腺癌是美国男性中癌症死亡（9％）的第二最常见的原因，旁边的肺和支气管（28％）1的癌症。根据最近的数据，估计是220，800初诊前列腺癌病例和27，540死亡将发生在2015年1。早期前列腺癌的五个年相对存活率为&gt; 99％，而晚期转移性疾病是只有28％1。治疗晚期转移性疾病的一个主要挑战是缺乏对本病的基本倾向的转移到其它器官，尤其是对骨骼，这是前列腺癌的一个频繁的部位分子机制的认识。因此，有一个明确的需要研究这些前列腺肿瘤的分子组成，以发展对发展有效的治疗方案，以先进的转移性疾病2,3。 前列腺肿瘤展览HIGħ生物异质没有明确定义的途径发展。转移往往与肿瘤的侵袭性4事先没有迹象发生。此临床异质性是由于前列腺癌的分子多样性。了解这些致命肿瘤的分子组成是设计出更好的诊断和治疗策略对本病的关键。因此，前列腺癌的研究目前主要集中在了解和预防转移。 临床前活体小鼠模型提供了多种选择，以了解前列腺癌的进展，以先进的转移性疾病的分子机制。此外，这些模型是对这种疾病的新的治疗策略的临床前评估重要。最常用的动物模型包括转基因小鼠模型，尾静脉注射，心内注入和人类原位小鼠模型。转基因研究是时间consumi纳克以及与人类的小鼠的前列腺癌的发展的相关性表明变性11。在自发转移小鼠模型，将细胞直接注射到循环和虽然，他们具有快速的周转时间，它们不能被用来研究在原发肿瘤或转移级联5的初始步骤。原位异种移植模型有显影骨转移灶，前列腺癌转移的常见部位的限制。然而，人类的原位前列腺癌异种移植的小鼠模型是充分表征的，并广泛用于研究原发性肿瘤发展，肿瘤和器官的微环境中，转移性疾病和使用的实验性药物用于治疗性干预6的初始相位之间的串扰的分子事件，7,8-11。

<table><tbody><tr><td>PC3 prostate cancer cell line&amp;nbsp;</td><td>ATCC</td><td>CRL-1435</td><td></td></tr><tr><td>Minimum Essential Medium (MEM)&amp;nbsp;</td><td>GIBCO,Life Technology</td><td>11095-080</td><td></td></tr><tr><td>PBS</td><td>GIBCO,Life Technology</td><td>10010-023</td><td></td></tr><tr><td>FBS</td><td>GIBCO,Life Technology</td><td>10437-028</td><td></td></tr><tr><td>Zeocin</td><td>Invitrogen,Life Technology</td><td>R250-01</td><td></td></tr><tr><td>Trypsin&amp;nbsp;</td><td>GIBCO,Life Technology</td><td>25300-54</td><td></td></tr><tr><td>IVIS&amp;nbsp;</td><td>Xenogen-Caliper</td><td></td><td></td></tr><tr><td>Insulin Syringes (300 &amp;micro;l, 28.5 g)</td><td>Becton Dickinson</td><td>309300</td><td></td></tr><tr><td>Mice</td><td>Charles River Laboratories, Inc</td><td></td><td></td></tr><tr><td>Alcohol Swabs</td><td>MEDEquip Depot</td><td>326895 BD</td><td></td></tr><tr><td>PVP Iodine Prep Pad</td><td>MEDEquip Depot</td><td>C12400PDI</td><td></td></tr><tr><td>Surgical CatGut Chromic Suture</td><td>Demetech</td><td>CC224017F0P</td><td></td></tr><tr><td>Matrigel</td><td>Corning</td><td>354248</td><td></td></tr></tbody></table>

pre-clinical orthotopic murine model of human prostate cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

以下PC3M吕克-C6细胞原位植入后前列腺叶，将小鼠每周用活的动物生物发光成像系统，以监测细胞的定居和肿瘤生长超过实验的过程中（ - B 图5A）成像。生物发光信号的定量分析表明，PC3M - 吕克 - C6细胞成功殖民前列腺叶。增加生物发光是指示增加原发性肿瘤生长在实验中（ 图5B）的过程中的。根据研究目的，小鼠可以每周非侵入性...

Watch this Scientific Journal Video about Pre-clinical Orthotopic Murine Model of Human Prostate Cancer at JoVE.com

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

人前列腺癌的临床前原位小鼠模型

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

pre-clinical-orthotopic-murine-model-of-human-prostate-cancer

Research

JoVE Journal

Medicine

14.2K 次观看.  VA Medical Center and UCSF. Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

视频: Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

A Murine Orthotopic Bladder Tumor Model and Tumor Detection System

This protocol describes the generation of bladder tumors in female C57BL/6J mice using the murine bladder cancer cell line MB49, which has been modified to secrete human Prostate Specific Antigen (PSA), and the procedure for the confirmation of tumor implantation. In brief, mice are anesthetized using injectable drugs and are made to lay in the dorsal position. Urine is vacated from the bladder and 50 &#181;L of poly-L-lysine (PLL) is slowly instilled at a rate of 10 &#181;L/20 s using a 24 G IV catheter. It is left in the bladder for 20 min by stoppering the catheter. The catheter is removed and PLL is vacated by gentle pressure on the bladder. This is followed by instillation of the murine bladder cancer cell line (1 x 105 cells/50 &#181;L) at a rate of 10 &#181;L/20 s. The catheter is stoppered to prevent premature evacuation. After 1 h, the mice are revived with a reversal drug, and the bladder is vacated. The slow instillation rate is important, as it reduces vesico-ureteral reflux, which can cause tumors to occur in the upper urinary tract and in the kidneys. The cell line should be well re-suspended to reduce clumping of cells, as this can lead to uneven tumor sizes after implantation.
This technique induces tumors with high efficiency. Tumor growth is monitored by urinary PSA secretion. PSA marker monitoring is more reliable than ultrasound or fluorescence imaging for the detection of the presence of tumors in the bladder. Tumors in mice generally reach a maximum size that negatively impacts health by about 3 - 4 weeks if left untreated. By monitoring tumor growth, it is possible to differentiate mice that were cured from those that were not successfully implanted with tumors. With only end-point analysis, the latter may be mistakenly assumed to have been cured by therapy.

This protocol describes the generation of murine orthotopic bladder tumors in female C57BL/6J mice and the monitoring of tumor growth.

小鼠原位膀胱肿瘤模型和肿瘤检测系统

This protocol describes the generation of bladder tumors in female C57BL/6J mice using the murine bladder cancer cell line MB49, which has been modified ...

科研

癌症研究

A Bioluminescent and Fluorescent Orthotopic Syngeneic Murine Model of Androgen-dependent and Castration-resistant Prostate Cancer

Orthotopic tumor modeling is a valuable tool for pre-clinical prostate cancer research, as it has multiple advantages over both subcutaneous and transgenic genetically engineered mouse models. Unlike subcutaneous tumors, orthotopic tumors contain more clinically accurate vasculature, tumor microenvironment, and responses to multiple therapies. In contrast to genetically engineered mouse models, orthotopic models can be performed with lower cost and in less time, involve the use of highly complex and heterogeneous mouse or human cancer cell lines, rather that single genetic alterations, and these cell lines can be genetically modified, such as to express imaging agents. Here, we present a protocol to surgically injecting a luciferase- and mCherry-expressing murine prostate cancer cell line into the anterior prostate lobe of mice. These mice developed orthotopic tumors that were non-invasively monitored in vivo and further analyzed for tumor volume, weight, mouse survival, and immune infiltration. Further, orthotopic tumor-bearing mice were surgically castrated, leading to immediate tumor regression and subsequent recurrence, representing castration-resistant prostate cancer. Although technical skill is required to carry out this procedure, this syngeneic orthotopic model of both androgen-dependent and castration-resistant prostate cancer is of great use to all investigators in the field.

The goal of this protocol is to demonstrate the intra-prostatic injection of prostate cancer cells, with subsequent castration. Orthotopic pre-clinical models of androgen-dependent and castration-resistant prostate cancer are critical to study the disease in the context of a clinically relevant tumor microenvironment and an immunocompetent host.

雄激素依赖性和去势抗性前列腺癌的荧光自体小鼠模型

Orthotopic tumor modeling is a valuable tool for pre-clinical prostate cancer research, as it has multiple advantages over both subcutaneous and ...

Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration

With an increasing incidence of prostate cancer, identification of new tumor drivers or modulators is crucial. Genetically engineered mouse models (GEMM) for prostate cancer are hampered by tumor heterogeneity and its complex microevolution dynamics. Traditional prostate cancer mouse models include, amongst others, germline and conditional knockouts, transgenic expression of oncogenes, and xenograft models. Generation of de novo mutations in these models is complex, time-consuming, and costly. In addition, most of traditional models target the majority of the prostate epithelium, whereas human prostate cancer is well known to evolve as an isolated event in only a small subset of cells. Valuable models need to simulate not only prostate cancer initiation, but also progression to advanced disease.
Here we describe a method to target a few cells in the prostate epithelium by transducing cells by viral particles. The delivery of an engineered virus to the murine prostate allows alteration of gene expression in the prostate epithelia. Virus type and quantity will hereby define the number of targeted cells for gene alteration by transducing a few cells for cancer initiation and many cells for gene therapy. Through surgery-based injection in the anterior lobe, distal from the urinary track, the tumor in this model can expand without impairing the urinary function of the animal. Furthermore, by targeting only a subset of prostate epithelial cells the technique enables clonal expansion of the tumor, and therefore mimics human tumor initiation, progression, as well as invasion through the basal membrane.
This novel technique provides a powerful prostate cancer model with improved physiological relevance. Animal suffering is limited, and since no additional breeding is required, overall animal count is reduced. At the same time, analysis of new candidate genes and pathways is accelerated, which in turn is more cost efficient.

This protocol describes a newly established method for virus delivery to the murine prostate. Using either CRISPR/Cas9 technology, gene overexpression, or Cre recombinase delivery, the technique allows orthotopic alteration of gene expression and implements a novel mouse model for prostate cancer.

CRISPR 指南对小鼠前列腺病毒传递的基因改变

With an increasing incidence of prostate cancer, identification of new tumor drivers or modulators is crucial. Genetically engineered mouse models (GEMM) ...

Identification, Histological Characterization, and Dissection of Mouse Prostate Lobes for In Vitro 3D Spheroid Culture Models

Genetically engineered mouse models (GEMMs) serve as effective pre-clinical models for investigating most types of human cancers, including prostate cancer (PCa). Understanding the anatomy and histology of the mouse prostate is important for the efficient use and proper characterization of such animal models. The mouse prostate has four distinct pairs of lobes, each with their own characteristics. This article demonstrates the proper method of dissection and identification of mouse prostate lobes for disease analysis. Post-dissection, the prostate cells can be further cultured in vitro for mechanistic understanding. Since mouse prostate primary cells tend to lose their normal characteristics when cultured in vitro, we outline here a method for isolating the cells and growing them as 3D spheroid cultures, which is effective for preserving the physiological characteristics of the cells. These 3D cultures can be used for analyzing cell morphology and behavior in near-physiological conditions, investigating altered levels and localizations of key proteins and pathways involved in the development and progression of a disease, and looking at responses to drug treatments.

Genetically engineered mice are useful models for investigating prostate cancer mechanisms. Here we present a protocol to identify and dissect prostate lobes from a mouse urogenital system, differentiate them based on histology, and isolate and culture the primary prostate cells in vitro as spheroids for downstream analyses.

体外3D 球形培养模型小鼠前列腺叶的鉴定、组织学特征及解剖

Genetically engineered mouse models (GEMMs) serve as effective pre-clinical models for investigating most types of human cancers, including prostate ...

MR Molecular Imaging of Prostate Cancer with a Small Molecular CLT1 Peptide Targeted Contrast Agent

Tumor extracellular matrix has abundance of cancer related proteins that can be used as biomarkers for cancer molecular imaging. In this work, we demonstrated effective MR cancer molecular imaging with a small molecular peptide targeted Gd-DOTA monoamide complex as a targeted MRI contrast agent specific to clotted plasma proteins in tumor stroma. We performed the experiment of evaluating the effectiveness of the agent for non-invasive detection of prostate tumor with MRI in a mouse orthotopic PC-3 prostate cancer model. The targeted contrast agent was effective to produce significant tumor contrast enhancement at a low dose of 0.03 mmol Gd/kg. The peptide targeted MRI contrast agent is promising for MR molecular imaging of prostate tumor.

To demonstrate MR cancer molecular imaging with a small peptide targeted MRI contrast agent specific to clotted plasma proteins in tumor stroma in a mouse prostate cancer model.

前列腺癌与小分子肽CLT1靶向造影剂的磁共振分子成像

Tumor extracellular  matrix has abundance of cancer related proteins that can be used as biomarkers  for cancer molecular imaging. In this work,  we ...

医学

Renal Capsule Xenografting and Subcutaneous Pellet Implantation for the Evaluation of Prostate Carcinogenesis and Benign Prostatic Hyperplasia

New therapies for two common prostate diseases, prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend critically on experiments evaluating their hormonal regulation. Sex steroid hormones (notably androgens and estrogens) are important in PrCa and BPH; we probe their respective roles in inducing prostate growth and carcinogenesis in mice with experiments using compressed hormone pellets. Hormone and/or drug pellets are easily manufactured with a pellet press, and surgically implanted into the subcutaneous tissue of the male mouse host. We also describe a protocol for the evaluation of hormonal carcinogenesis by combining subcutaneous hormone pellet implantation with xenografting of prostate cell recombinants under the renal capsule of immunocompromised mice. Moreover, subcutaneous hormone pellet implantation, in combination with renal capsule xenografting of BPH tissue, is useful to better understand hormonal regulation of benign prostate growth, and to test new therapies targeting sex steroid hormone pathways.

We describe the manufacture of compressed hormone pellets, as well as subcutaneous surgical implantation into mice. This strategy can be combined with the growth of cell and tissue xenografts under the renal capsule of athymic nude mice to evaluate hormonal carcinogenesis and regulation of benign prostate growth.

肾小囊和异种移植皮下颗粒植入前列腺癌发生和良性前列腺增生症的评价

New therapies for two common prostate diseases,  prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend  critically on experiments ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

人类前列腺癌转移的原位小鼠模型

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Intra-Cardiac Injection of Human Prostate Cancer Cells to Create a Bone Metastasis Xenograft Mouse Model

As the most common male malignancy, prostate cancer (PC) ranks second in mortality, primarily due to a 65%-75% bone metastasis rate. Therefore, it is essential to understand the process and related mechanisms of prostate cancer bone metastasis for developing new therapeutics. For this, an animal model of bone metastasis is an essential tool. Here, we report detailed procedures to generate a bone metastasis mouse model via intra-cardiac injection of prostate cancer cells. A bioluminescence imaging system can determine whether prostate cancer cells have been accurately injected into the heart and monitor cancer cell metastasis since it has great advantages in monitoring metastatic lesion development. This model replicates the natural development of disseminated cancer cells to form micro-metastases in the bone and imitates the pathological process of prostate cancer bone metastasis. It provides an effective tool for further exploration of the molecular mechanisms and the in vivo therapeutic effects of this disease.

Here, we present a protocol for the intra-cardiac injection of human prostate cancer cells to generate a mouse model with bone metastasis lesions.

心内注射人前列腺癌细胞以创建骨转移异种移植小鼠模型

As the most common male malignancy, prostate cancer (PC) ranks second in mortality, primarily due to a 65%-75% bone metastasis rate. Therefore, it is ...

Generation of Prostate Cancer Patient Derived Xenograft Models from Circulating Tumor Cells

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, biologic studies, and personalized medicine strategies. Circulating tumor cells (CTC) play a critical role in tumor metastasis and have been isolated from patients with several tumor types. Recently, CTCs have been used to generate PDX experimental models of breast and prostate cancer. This manuscript details the method for the generation of prostate cancer PDX models from CTCs developed by our group. Advantages of this method over conventional PDX models include independence from surgical sample collection and generating experimental models at various disease stages. Density gradient centrifugation followed by red blood cell lysis and flow cytometry depletion of CD45 positive mononuclear cells is used to enrich CTCs from peripheral blood samples collected from patients with metastatic disease. The CTCs are then injected into immunocompromised mice; subsequently generated xenografts can be used for functional studies or harvested for molecular characterization. The primary limitation of this method is the negative selection method used for CTC enrichment. Despite this limitation, the generation of PDX models from CTCs provides a novel experimental model to be applied to prostate cancer research.

This manuscript details a method used to generate prostate cancer patient derived xenografts (PDXs) from circulating tumor cells (CTCs). The generation of PDX models from CTCs provides an alternative experimental model to study prostate cancer; the most commonly diagnosed tumor and a frequent cause of death from cancer in men.

前列腺癌患者产生派生异种移植模型从循环肿瘤细胞

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, ...

Murine Prostate Micro-dissection and Surgical Castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse. We also depict representative results produced by these protocols. Finally, we discuss the advantages and utilization of these protocols.

鼠前列腺微解剖和手术去势

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...