3D培养在神经胶质瘤研究中的应用

传统的肿瘤研究依赖于肿瘤细胞系构建肿瘤模型,并在细胞基础上测定肿瘤细胞的耐药性等,但在临床实验中大多数肿瘤药物以失败告终,其中最主要的原因是细胞系不能模拟肿瘤细胞在人体内微环境的特性,3D培养技术在肿瘤研究中的应用愈加广泛。本文主要从3D培养在神经胶质瘤研究中的应用进行综述。     

3D打印技术在蝶骨嵴脑膜瘤切除术中的应用价值

  目的  研究3D打印技术在蝶骨嵴脑膜瘤手术中的应用价值。  方法  选取福建省立医院2016年9月至2016年12月经头颅MRI及螺旋CT扫描发现的蝶骨嵴脑膜瘤患者5例，利用颅脑螺旋CT增强扫描的DICOM图像，针对颅骨、血管及肿瘤，分别进行数据信息的提取和重建，并在同一坐标系下实现装配融合，建立三维复合虚拟模型，再经3D打印技术制作出3D实体模型，于术前、术中参考。  结果  成功重建5例患者的蝶骨嵴脑膜瘤三维复合虚拟模型，并制作出实体解剖模型，模型清晰显示颅骨、肿瘤及其与毗邻血管的位置关系，将其应用于术前规划与术中参考，5例患者手术均获得成功。  结论  基于3D打印技术的蝶骨嵴脑膜瘤三维复合虚拟模型及其实体解剖模型在优化手术预案、提高肿瘤切除率、减少手术损伤等方面具有重要意义。      

肿瘤模型研究进展及应用

肿瘤模型研究是肿瘤领域的热点。肿瘤模型为肿瘤机制研究及临床转化应用提供了基础保障。肿瘤细胞系和异种移植物是目前常用的研究模型,但因细胞系缺乏原始肿瘤的异质性,异种移植物培养周期长、成瘤率低,应用存在一定局限性。肿瘤3D培养模型包括肿瘤悬浮球和肿瘤类器官模型,模拟了原始肿瘤的生长环境,且很好地模拟了细胞间及细胞-基质间的相互作用。肿瘤类器官作为新兴的肿瘤研究平台,具有易于构建、成瘤率高,并能长期传代和冷冻保存等优点,在很大程度上维持原始肿瘤的结构、遗传特性和肿瘤内异质性,在肿瘤研究中具有巨大潜力。      

类器官在肿瘤转化医学中的应用和进展

随着精准医学概念的提出，肿瘤作为一类高度异质性的疾病，其个体化治疗已成为精准医学的一个关键领域也受到更多关注。近期提出的类器官模型为肿瘤的基础研究和个体化治疗带来了新突破。类器官指从干细胞或器官祖细胞来源，以类似体内细胞分化的方式组织成的器官特异性的细胞集合。肿瘤类器官指利用原代恶性细胞经过体外3D培养构建的恶性细胞团，在体外培养条件下可持续增殖，一定程度上保留了原代肿瘤的病理学形态特征、基因组与转录组特征、药物敏感性及恶性细胞间异质性，为体外肿瘤研究提供了新方法，尤其在预测患者药物敏感性、药物高通量筛选等方面有巨大潜力，为肿瘤个体化治疗作出了贡献。但该模型仍存在不足，如无法重现体内肿瘤微环境等，使其在临床等方面的应用受限。目前将肿瘤类器官与其他肿瘤成分共培养、与微流控设备和生物打印技术等联合应用将有望弥补其缺陷、突破肿瘤治疗领域的瓶颈。本文将常用肿瘤研究模型的特点进行对比，总结部分肿瘤类器官的培养方法，并描述了肿瘤类器官的临床相关应用，最后对其与其他技术联合应用进行了叙述，并对未来肿瘤类器官的发展方向予以展望。     

3D打印技术在骨肿瘤手术应用中的研究进展

 随着影像图像后处理技术及3D打印技术的快速发展，保肢手术成为骨肿瘤患者的首要选择，而彻底完整切除肿瘤、骨缺损和重建患肢功能是面临的主要挑战。3D打印是一种快速成型技术，可实现解剖模型、导航模板、肿瘤假体的个体化制造以及骨组织工程生物3D打印等，被广泛应用于骨肿瘤的保肢治疗且取得了理想的临床效果。本文就3D打印技术在骨肿瘤个体化修复重建应用中的研究进展进行综述。     

Matrigel三维培养模型对乳腺癌细胞整合素β１的定位及胞外基质粘附的影响

目的：观察Matrigel三维（3D）培养模型对乳腺癌细胞形貌、细胞整合素β1的定位及其介导的细胞与胞外基质（ECM）层粘连蛋白,纤连蛋白和Ⅳ型胶原蛋白粘附能力的影响。初步探索在二维（2D）和3D不同培养模式下乳腺癌细胞与ECM的相互作用,揭示肿瘤微环境对肿瘤细胞的影响。方法：分别在2D及3D培养模型下进行乳腺癌细胞MDA-MB-435S及MDA-MB-231的体外培养,钙黄绿素Calcein AM检测细胞存活力,免疫荧光分析整合素β1的定位,粘附率实验检测细胞对ECM各成分的粘附变化。结果：整合素β1在2D及3D培养模型中的定位具有显著差异,其介导的细胞与层粘连蛋白的黏附能力在3D模型中显著下降,而细胞与纤连蛋白和Ⅳ型胶原蛋白的粘附能力明显上升。结论：与2D培养结果不同,3D培养模型更能模拟体内3D微环境,可做为新的研究模式揭示肿瘤细胞与ECM的相互作用,以及肿瘤微环境在肿瘤进展中的作用。     

基于microcarrier 6构建正常免疫小鼠人胃癌移植模型

  目的   基于新型3D微载体（microcarrier 6）复合人胃癌MKN45细胞接种于具有正常免疫功能的小鼠，以期建立新型人胃癌正常小鼠转移瘤模型   方法   将60只雄性C57BL/6小鼠按是否将MKN45细胞接种于支架材料随机分为2D组、对照组和3D组；体外构建三维肿瘤细胞培养模型，采用皮下接种法建立小鼠移植瘤模型，观察小鼠成瘤时间、成瘤率、病理学表现等。   结果   2D组和对照组均未成瘤，3D组成瘤率达80%，成瘤时间早，移植瘤HE染色和免疫组织化学均符合人胃癌特点。   结论   本实验成功基于microcarrier 6复合人胃癌MKN45细胞在正常免疫小鼠中建立人胃癌小鼠移植瘤模型，此模型可以更好地研究和进一步阐明胃癌在免疫功能正常机体中发生、发展机制，同时也为抗癌药物的研发提供了更有价值的动物模型。      

维生素 D 3 与胃肠道肿瘤的研究进展

维生素D3结构上属于甾体激素,它的主要生理功能是调节钙磷代谢和维持正常血钙水平、调节骨代谢和多种细胞的生理反应。近年来很多研究表明维生素D3在多种肿瘤中扮演重要角色,影响肿瘤细胞的增殖、凋亡等活动。维生素D3作用于不同癌细胞类型的细胞机制,有力地表明维生素D3可以发挥保护和抗肿瘤作用,延缓细胞转化、增生和肿瘤进展。流行病学研究发现血清维生素D水平与胃肠道肿瘤发病风险呈负相关,血清维生素D水平与胃肠道肿瘤患者预后正相关。进一步研究表明维生素D3可以通过多种途径诱导细胞凋亡、阻滞细胞周期、抑制胃肠道肿瘤细胞增殖。维生素D3能够与化疗药物协同作用促进肿瘤细胞凋亡、抑制肿瘤细胞增殖。维生素D3类似物不产生高血钙的副作用,也能够发挥其抑制肿瘤细胞生长的作用。维生素D3可能通过维生素D3受体VDR发挥其抗肿瘤作用。结合这些研究表明维生素D3有希望做为治疗和预防胃肠道肿瘤的新思路,更多的维生素D3抗肿瘤机制研究及临床实验能够为胃肠道肿瘤患者带来福音。     

Cyclin D3与肿瘤的研究进展

细胞周期的失调在肿瘤的发生发展过程中起着关键性作用。细胞周期蛋白D(Cyclin D)是G1/S期转换的重要正性调控因子，包括3个亚型：Cyelin D1、Cyclin D2和Cyelin D3。已证实Cyelin D1是1种原癌基因，其基因结构和功能异常与多种肿瘤(如乳腺癌、食管癌、肺癌、卵巢癌、黑色素瘤等)的发生发展密切相关。Cyclin D2在胃癌、结直肠癌、骨肉瘤、慢性B细胞淋巴细胞白血病等肿瘤中发现有异常表达。     

3D打印模型辅助肩胛骨软骨肉瘤的地图形外科切除CSCD

目的描述一种地图形切除肩胛骨肿瘤的手术方法,并评估其切除精确性、肿瘤学预后和肩关节功能。方法回顾分析北京积水潭医院2013-2017年6例因肩胛骨肿瘤接受了3D打印模型辅助地图形外科切除手术患者的资料。术前采用CT扫描获得肿瘤数据。将图像数据集导入导航系统(Stryker Navigation),对肿瘤进行标记,术前设计截骨边缘。根据标有肿瘤信息的图像数据打印3D模型。术中按3D打印模型和术前设计截骨线完成手术。将术前计划和实际获得的切缘作比较。对外科边界、肿瘤局部复发、远处转移及肩关节功能进行评估。结果术前计划切缘21.0±8.1 mm,实际手术获得切缘22.0±6.6mm,两者匹配差值为1.0±5.3mm,差异无统计学意义(t=-0.795,P=0.437)。平均随访51.6月,未发现局部复发和远处转移,Neer肩关节功能评分96~100分,平均为98.2分,MSTS功能评分97%~100%,平均99%。结论使用3D打印模型辅助肩胛骨软骨肉瘤的地图形外科切除可以获得精确的肩胛骨部分切除及理想的肿瘤控制效果,术后肩关节功能满意。     

Spherical Cancer Models in Tumor Biology

Three-dimensional (3D) in vitro models have been used in cancer research as an intermediate model between in vitro cancer cell line cultures and in vivo tumor. Spherical cancer models represent major 3D in vitro models that have been described over the past 4 decades. These models have gained popularity in cancer stem cell research using tumorospheres. Thus, it is crucial to define and clarify the different spherical cancer models thus far described. Here, we focus on in vitro multicellular spheres used in cancer research. All these spherelike structures are characterized by their well-rounded shape, the presence of cancer cells, and their capacity to be maintained as free-floating cultures. We propose a rational classification of the four most commonly used spherical cancer models in cancer research based on culture methods for obtaining them and on subsequent differences in sphere biology: the multicellular tumor spheroid model, first described in the early 70s and obtained by culture of cancer cell lines under nonadherent conditions; tumorospheres, a model of cancer stem cell expansion established in a serum-free medium supplemented with growth factors; tissue-derived tumor spheres and organotypic multicellular spheroids, obtained by tumor tissue mechanical dissociation and cutting. In addition, we describe their applications to and interest in cancer research; in particular, we describe their contribution to chemoresistance, radioresistance, tumorigenicity, and invasion and migration studies. Although these models share a common 3D conformation, each displays its own intrinsic properties. Therefore, the most relevant spherical cancer model must be carefully selected, as a function of the study aim and cancer type.     

字化重建人体立体X线透视图研究

本文给出应用于肿瘤放射治疗领域的基于CT影像序列的数字化重建人体立体X线透视图（three dimensional digitally reconstructed radiograph，3D DRR）研究结果，包括实现3D DRR技术的简要步骤及原理，以及临床应用实例。借助于3D DRR技术，不仅可以准确地确定肿瘤及周围正常组织器官的空间形态关系，而且可以准确地验证放射治疗中各方向射柬照射肿瘤靶区的空间适形度，以及周围正常组织器官是否受到了不利的照射影响。     

In vitro three‐dimensional (3D) models in cancer research: An update

Tissues are three‐dimensional (3D) entities as is the tumor that arises within them. Though disaggregated cancerous tissues have produced numerous cell lines for basic and applied research, it is generally agreed that these lines are poor models of in vivo phenomena. In this review we focus on in vitro 3D models used in cancer research, particularly their contribution to molecular studies of the early stages of metastasis, angiogenesis, the tumor microenvironment, and cancer stem cells. We present a summary of the various formats used in the field of tissue bioengineering as they apply to mechanistic modeling of cancer stages or processes. In addition we list studies that model specific types of malignancies, highlight drastic differences in results between 3D in vitro models and classical monolayer culturing techniques, and establish the need for standardization of 3D models for meaningful preclinical and therapeutic testing. © 2011 Wiley Periodicals, Inc.     

Action of low calcemic 1alpha,25-dihydroxyvitamin D3 analogue EB1089 in head and neck squamous cell carcinoma

BACKGROUND: 1alpha,25-Dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] and its analogues inhibit growth of various types of cancer cells. Although the therapeutic potential of 1,25(OH)(2)D(3) is limited by its tendency to induce hypercalcemia, analogues such as EB1089 are potent inhibitors of cell growth and exhibit reduced calcemic effects. We analyzed the antiproliferative and calcemic effects of EB1089 in tissue culture and animal models of head and neck squamous cell carcinoma (SCC) to investigate its potential as a chemotherapeutic/chemopreventive agent. METHODS: The effects of 1,25(OH)(2)D(3) and EB1089 on cell growth and expression of p21(WAF1/CIP1) and p27(KIP1), which encode cyclin-dependent kinase inhibitors, and a novel target, gadd45alpha, a growth-arrest and DNA-damage gene, were monitored in cultured murine AT-84 SCC cells. The effects of these agents on AT-84 cell growth in vitro and on growth of AT-84 tumors in syngeneic C3H mice were monitored; treatment started at the time of tumor implantation (early tumor model) or after 12 days (late tumor model). Weight and serum calcium levels were also monitored in these animals. All P values were two-sided. RESULTS: Both 1,25(OH)(2)D(3) and EB1089 arrested proliferation of AT-84 cells in G(0)/G(1) phase, inhibited p21(WAF1/CIP1) expression, and induced expression of p27(KIP1) protein. 1,25(OH)(2)D(3) also enhanced the expression of gadd45alpha, apparently by a p53-independent mechanism. There was a statistically significant decrease in tumor growth for 1,25(OH)(2)D(3)-treated mice (P<.001 for early tumor model) and EB1089-treated mice (P<.001 and P =.001 for early and late tumor models, respectively). Unlike 1,25(OH)(2)D(3), EB1089 did not induce cachexia or hypercalcemia. The effects of 1,25(OH)(2)D(3) and EB1089 on expression of p21(WAF1/CIP1) and GADD45alpha were similar in tumors and in vitro. CONCLUSIONS: EB1089 completely inhibited growth of AT-84 SCC cells at nanomolar concentrations, reduced tumor growth, and did not have calcemic effects. Our results support continued investigation of EB1089 as a chemopreventive/chemotherapeutic agent for head and neck SCC.     

A vitamin D receptor-alkylating derivative of 1α,25-dihydroxyvitamin D3 inhibits growth of human kidney cancer cells and suppresses tumor growth

1,25-Dihydroxyvitamin D? [1,25(OH)?D?] has shown strong promise as an antiproliferative agent in several malignancies, yet its therapeutic use has been limited by its toxicity leading to search for analogues with antitumor property and low toxicity. In this study, we evaluated the in vitro and in vivo properties of 1,25-dihydroxyvitamin D?-3-bromoacetate [1,25(OH)?D?-3-BE], an alkylating derivative of 1,25(OH)?D?, as a potential therapeutic agent for renal cancer. Dose response of 1,25(OH)?D?-3-BE in 2 kidney cancer cell lines was evaluated for its antiproliferative and apoptotic properties, and mechanisms were evaluated by Western blot and FACS analyses. Therapeutic potential of 1,25(OH)?D?-3-BE was assessed both by determining its stability in human serum and by evaluating its efficacy in a mouse xenograft model of human renal tumor. We observed that 1,25(OH)?D?-3-BE is significantly more potent than an equivalent concentration of 1,25(OH)?D? in inhibiting growth of A498 and Caki 1 human kidney cancer cells. 1,25(OH)?D?-3-BE-mediated growth inhibition was promoted through inhibition of cell-cycle progression by downregulating cyclin A and induction of apoptosis by stimulating caspase activity. Moreover, 1,25(OH)?D?-3-BE strongly inhibited Akt phosphorylation and phosphorylation of its downstream target, caspase-9. 1,25(OH)?D?-3-BE seemed to be stable in human serum. In xenograft mouse model of human renal tumor, 1,25(OH)?D?-3-BE was more potent at reducing tumor size than 1,25(OH)?D?, which was accompanied by an increase in apopotosis and reduction of cyclin A staining in the tumors. These results suggest a translational potential of this compound as a therapeutic agent in renal cell carcinoma. Data from this study and extensive studies of vitamin D for the prevention of many malignancies support the potential of 1,25(OH)?D?-3-BE for preventing renal cancer and the development of relevant in vivo prevention models for assessing this potential, which do not exist at present.     

D-二聚体与转移癌的关系及在口腔肿瘤中的展望

血浆D-二聚体是交联纤维蛋白水解后的特异性产物,其水平升高反映了机体凝血异常存在继发性纤溶亢进。恶性肿瘤患者血液往往处于高凝状态,这种状态有利于肿瘤细胞逃避机体免疫和扩散转移。近几年来发现血浆D-二聚体水平与多种恶性肿瘤的分期和转移密切相关,可作为肿瘤转移的预测因子。本文就血浆D-二聚体在转移癌中的研究情况作一综述,并对在口腔颌面部恶性肿瘤中的应用可能作一展望。     

Biomaterial-based platforms for cancer stem cell enrichment and study

Cancer stem cells (CSCs) are a relatively rare subpopulation of tumor cell with self-renewal and tumorigenesis capabilities. CSCs are associated with cancer recurrence, progression, and chemoradiotherapy resistance. Establishing a reliable platform for CSC enrichment and study is a prerequisite for understanding the characteristics of CSCs and discovering CSC-related therapeutic strategies. Certain strategies for CSC enrichment have been used in laboratory, particularly fluorescence-activated cell sorting (FACS) and mammosphere culture. However, these methods fail to recapitulate the in vivo chemical and physical conditions in tumors, thus potentially decreasing the malignancy of CSCs in culture and yielding unreliable research results. Accumulating research suggests the promise of a biomaterial-based three-dimensional (3D) strategy for CSC enrichment and study. This strategy has an advantage over conventional methods in simulating the tumor microenvironment, thus providing a more effective and predictive model for CSC laboratory research. In this review, we first briefly discuss the conventional methods for CSC enrichment and study. We then summarize the latest advances and challenges in biomaterial-based 3D CSC platforms. Design strategies for materials, morphology, and chemical and physical cues are highlighted to provide direction for the future construction of platforms for CSC enrichment and study.     

Volume Reconstruction Techniques Improve the Correlation Between Histological and in vivo Tumor Volume Measurements in Mouse Models of Human Gliomas

Assessment of therapy efficacy using animal models of tumorigenic cancer requires the ability to accurately measure changes in tumor volume over the duration of disease course. In order to be meaningful, in vivo tumor volume measurements by non-invasive techniques must correlate with tumor volume measurements from endpoint histological analysis. Tumor volume is frequently assessed by endpoint histological analyses approximating the tumor volume with geometric primitives such as spheroids and ellipsoids. In this study we investigated alternative techniques for quantifying histological volume measurements of tumors in a xenograft orthotopic mouse model of human glioblastoma multiforme, and compared these to in vivo tumor volume measurements based on magnetic resonance imaging (MRI) data. Two techniques leveraging three-dimensional (3D) image analysis methods were investigated. The first technique involves the reconstruction of a smoothed polygonal model representing the tumor volume from histological section images and is intended for accuracy and qualitative assessment of tumor burden by visualization, while a second technique which approximates the tumor volume as a series of slabs is presented as an abbreviated process intended to produce quantitatively similar volume measurements with a minimum of effort required on behalf of the investigator. New software (QuickVol) designed for use in the first technique, is also discussed. In cases where tumor growth is asymmetric and invasive, we found that 3D analysis techniques using histological section images produced volume measurements more consistent with in vivo volume measurements based on MRI data, than approximation of tumor volume using geometric primitives. Visualizations of the volumes represented by each of these techniques qualitatively support this finding, and suggest that future research using mouse models of glioblastoma multiforme (genetically engineered or xenograft) will benefit from the use of these or similar alternative tumor volume measurement techniques.