A biomimetic nanoplatform for customized photothermal therapy of HNSCC evaluated on patient-derived xenograft models

Cancer cell membrane(CCM)derived nanotechnology functionalizes nanoparticles(NPs)to recognize homologous cells,exhibiting translational potential in accurate tumor therapy.However,these nanoplatforms are majorly generated from fixed cell lines and are typically evaluated in cell line-derived subcutaneous-xenografts(CDX),ignoring the tumor heterogeneity and differentiation from inter-and intra-individuals and microenvironments between heterotopic-and orthotopic-tumors,limiting the therapeutic efficiency of such nanoplatforms.Herein,various biomimetic nanoplatforms(CCM-modified gold@Carbon,i.e.,Au@C-CCM)were fabricated by coating CCMs of head and neck squamous cell carcinoma(HNSCC)cell lines and patient-derived cells on the surface of Au@C NP.The generated Au@C-CCMs were evaluated on corresponding CDX,tongue orthotopic xenograft(TOX),immunecompetent primary and distant tumor models,and patient-derived xenograft(PDX)models.The Au@C-CCM generates a photothermal conversion efficiency up to 44.2% for primary HNSCC therapy and induced immunotherapy to inhibit metastasis via photothermal therapy-induced immunogenic cell death.The homologous CCM endowed the nanoplatforms with optimal targeting properties for the highest therapeutic efficiency,far above those with mismatched CCMs,resulting in distinct tumor ablation and tumor growth inhibition in all four models.This work reinforces the feasibility of biomimetic NPs combining modular designed CMs and functional cores for customized treatment of HNSCC,can be further extended to other malignant tumors therapy.

Translational pancreatic cancer research:a comparative study on patient-derived xenograft models

AIM To assess the viability of orthotopic and heterotopic patient-derived pancreatic cancer xenografts implanted into nude mice.METHODS This study presents a prospective experimental analytical follow-up of the development of tumours in mice upon implantation of human pancreatic adenocarcinoma samples. Specimens were obtained surgically from patients with a pathological diagnosis of pancreatic adenocarcinoma. Tumour samples from pancreatic cancer patients were transplanted into nude mice in three different locations(intraperitoneal, subcutaneous and pancreatic). Histological analysis(haematoxylin-eosin and Masson's trichrome staining) and immunohistochemical assessment of apoptosis(TUNEL), proliferation(Ki-67), angiogenesis(CD31) and fibrogenesis(α-SMA) were performed. When a tumour xenograft reached the target size, it was reimplanted in a new nude mouse. Three sequential tumour xenograft generations were generated(F1, F2 and F3).RESULTS The overall tumour engraftment rate was 61.1%. The subcutaneous model was most effective in terms of tissue growth(69.9%), followed by intraperitoneal(57.6%) and pancreatic(55%) models. Tumour development was faster in the subcutaneous model(17.7 ± 2.6 wk) compared with the pancreatic(23.1 ± 2.3 wk) and intraperitoneal(25.0 ± 2.7 wk) models(P = 0.064). There was a progressive increase in the tumour engraftment rate over successive generations for all three models(F1 28.1% vs F2 71.4% vs F3 80.9%, P < 0.001). There were no significant differences in tumour xenograft differentiation and cell proliferation between human samples and the three experimental models among the sequential generations of tumour xenografts. However, a progressive decrease in fibrosis, fibrogenesis, tumour vascularisation and apoptosis was observed in the three experimental models compared with the human samples. All three pancreatic patient-derived xenograft models presented similar histological and immunohistochemical characteristics.CONCLUSION In our experience, the faster development andgreatest number of viable xenografts could make the subcutaneous model the best option for experimentation in pancreatic cancer.

Advantages and limitations in the establishment and utilization of patient-derived xenografts in gastric cancer

Owing to the high genetic heterogeneity of tumors, small number of therapeutic strategies available, and frequent presentation of drug resistance, the prognosis for patients with advanced gastric cancer(AGC) are unsatisfactory. The utility of traditional cancer cell lines in translational research is limited by their poor correspondence to the genomic alterations and expression profiles that occur in actual patient tumors. In the last decade, increasing attention has been given to patient-derived tumor xenografts(PDTXs), which can faithfully recapitulate the histopathology, molecular characteristics, and therapeutic responses of the patient's tumor. However, the widespread development and utilization of PDTXs is restricted by factors such as the timeframe of establishment, lymphoma transformation during passaging, the immunodeficient microenvironment, and pharmacokinetic differences between mice and humans. In this review, we summarize the establishment and characterization of PDTX models for gastric cancer(GC). We then weigh the advantages and limitations of PDTXs when used to evaluate novel compounds, identify effective biomarkers, demonstrate resistance mechanisms, and predict clinical outcomes.

The promises and challenges of patient-derived tumor organoids in drug development and precision oncology

In the era of precision medicine,cancer researchers and oncologists are eagerly searching for more realistic,cost effective,and timely tumor models to aid drug development and precision oncology.Tumor models that can faithfully recapitulate the histological and molecular characteristics of various human tumors will be extremely valuable in increasing the successful rate of oncology drug development and discovering the most efficacious treatment regimen for cancer patients.Two‐dimensional(2D)cultured cancer cell lines,genetically engineered mouse tumor(GEMT)models,and patient‐derived tumor xenograft(PDTX)models have been widely used to investigate the biology of various types of cancers and test the efficacy of oncology drug candidates.However,due to either the failure to faithfully recapitulate the complexity of patient tumors in the case of 2D cultured cancer cells,or high cost and untimely for drug screening and testing in the case of GEMT and PDTX,new tumor models are urgently needed.The recently developed patient‐derived tumor organoids(PDTO)offer great potentials in uncovering novel biology of cancer development,accelerating the discovery of oncology drugs,and individualizing the treatment of cancers.In this review,we will summarize the recent progress in utilizing PDTO for oncology drug discovery.In addition,we will discuss the potentials and limitations of the current PDTO tumor models.

Quantitative examination of the inhibitory activation of molecular targeting agents in hepatocellular carcinoma patient-derived cell invasion via a novel in vivo tumor model

Background: The outcomes for patients with advanced hepatocellular carcinoma(HCC) receiving sorafenib are far from satisfactory because of treatment resistance to sorafenib. However, the exact mechanism of resistance to sorafenib remains unclear and it is valuable to establish a novel mouse model to quantitatively analyze the inhibition rates of sorafenib on the invasive growth of HCC cells in the liver.Methods: HCC tissue microblocks derived from patients were cultured and mixed with hydrogel drops. Then, hydrogel drops containing microblocks of HCC tissue were attached onto the surface of the livers of nude mice to form lesions or nodules of HCC. The mice received molecular targeting agents through oral administration. Livers with tumor nodules were harvested for H&E staining(hematoxylin-eosin staining) analysis and H&E staining images were quantitatively analyzed using image J software. The invasive growth of HCC cells into the liver was calculated using the depth of the lesions compared with the total thickness of the liver.Results: Microblocks containing cells derived from HCC patients can form lesions in the liver of nude mice. Oral administration of molecular targeting agents inhibited the invasive growth of HCC cells in the liver of nude mice.Conclusions: The model established in this study involves the invasive growth of HCC cells in the liver of nude mice, and the model allows for the quantitative analysis of the inhibitory effect of molecular targeting agents on the invasion of HCC cells in vivo.

Establishment,functional and genetic characterization of a colon derived large cell neuroendocrine carcinoma cell line

AIM To establish cell line and patient-derived xenograft(PDX) models for neuroendocrine carcinomas(NEC) which is highly desirable for gaining insight into tumor development as well as preclinical research includingbiomarker testing and drug response prediction.METHODS Cell line establishment was conducted from direct in vitro culturing of colonic NEC tissue(HROC57). A PDX could also successfully be established from vitally frozen tumor samples. Morphological features, invasive and migratory behavior of the HROC57 cells as well as expression of neuroendocrine markers were vastly analyzed. Phenotypic analysis was done by microscopy and multicolor flow cytometry. The extensive molecular-pathological profiling included mutation analysis, assessment of chromosomal and microsatellite instability; and in addition, fingerprinting(i.e., STR analysis) was performed from the cell line in direct comparison to primary patient-derived tissues and the PDX model established. Drug responsiveness was examined for a panel of chemotherapeutics in clinical use for the treatment of solid cancers.RESULTS The established cell line HROC57 showed distinct morphological and molecular features of a poorly differentiated large-cell NEC with KI-67 > 50%. Molecular-pathological analysis revealed a Cp G island promoter methylation positive cell line with microsatellite instability being absent. The following mutation profile was observed: KRAS(wt), BRAF(mut). A high sensitivity to etoposide, cisplatin and 5-FU could be demonstrated while it was more resistant towards rapamycin. CONCLUSION We successfully established and characterized a novel patient-derived NEC cell line in parallel to a PDX model as a useful tool for further analysis of the biological characteristics and for development of novel diagnostic and therapeutic options for NEC.

头颈部恶性肿瘤研究模型的演化

恶性肿瘤发生发展的机制探索以及抗癌药物治疗疗效的评估均有赖于各种体内与体外研究模型的建立。近几十年间,随着生物医学技术的快速发展,恶性肿瘤的体内外研究模型也发生了巨大的变化。基因检测技术从单基因到多基因的进展促进了生物信息学飞速发展和恶性肿瘤概念的转变;体外细胞研究模型从单层的二维培养、原代培养向立体的三维构型发展,从而更好地重现肿瘤组织的细胞间交互作用与功能;体内动物研究模型由传统的致癌物诱导、细胞或组织形成移植瘤逐渐演变为基因编辑的动物模型或人源性肿瘤异种移植模型,从而可以针对性地研究相关基因在肿瘤发生发展中的作用;传统的临床研究也从简单的临床回顾性研究更多地向前瞻性研究转变,Ⅰ期/Ⅱ期/Ⅲ期临床研究,研究者发起的临床研究以及真实世界临床研究,这些研究为临床研究增添了活力。目前恶性肿瘤研究模型存在的主要不足包括模型的单一性、对肿瘤微环境的模拟不足、动物肿瘤模型与人类肿瘤差异性,以及缺乏对个性化医疗的考量。未来仍需要进一步研发和优化研究模型,并更有效地将不同模型整合起来,形成一个优化的整体实验模型系统。本文将系统回顾恶性肿瘤研究模型的演化并对相关模型进行阐述,为科研工作者进行恶性肿瘤的研究提供合理的研究模型。

人源性胰母细胞瘤异种移植模型的构建与验证

目的研究构建合适的动物模型用于胰母细胞瘤(PBL)的化疗药物筛选及新药评估。方法将手术切除的PBL肿瘤组织碎块移植到免疫缺陷的小鼠皮下组织,待移植瘤生长至1500 mm^(3)时进行瘤体传代及保存。使用HE染色、免疫组织化学及PCR法对成功建立的PBL患儿起源的异种移植(PDX)模型进行人源性鉴定。结果HE染色显示小鼠PDX中的肿瘤组织及细胞和患儿肿瘤细胞形态基本相同,免疫组化结果显示β-连环蛋白(β-Catenin)、细胞角蛋白7(CK7)和细胞角蛋白19(CK19)在PDX模型组织与人属源性肿瘤组织中有相同的免疫学特征。PCR分析显示PDX组织和原发肿瘤组织具有相同的个体来源。结论所建立胰母细胞瘤来源PDX高度保留了原发胰母细胞瘤的生长模式及蛋白表达特性,为胰母细胞瘤患者的基础研究和临床用药指导提供了重要的实验模型工具,给推动疾病机制探索及制定个性化治疗方案提供了科学依据。

肺癌脑转移瘤人源异种移植模型的构建及应用研究进展

肺癌脑转移瘤是中枢神经系统最为常见的继发性恶性肿瘤,目前共识的治疗方式是在安全范围内通过手术最大程度切除肿瘤组织,术后进行化学治疗以及局部或全身放射治疗,但其治疗效果仍不理想。人源异种移植瘤模型能够很好地保留患者原发肿瘤异质性、组织学特性和分子多样性,能够为研究新的治疗方法提供新思路及新的研究工具。本文就移植瘤模型的建立、模型在肺癌脑转移瘤治疗中的应用研究以及模型的应用前景和局限性进行综述。

肺癌恶性胸腔积液来源肿瘤细胞的小鼠PDX模型构建及实验验证

目的·构建肺癌患者恶性胸腔积液(malignant pleural effusion,MPE)肿瘤细胞来源的肿瘤异种移植(patientderived tumor xenograft,PDX)模型,并进行实验验证。方法·从基因表达综合数据集(Gene Expression Omnibus,GEO)下载人肺癌伴MPE单细胞转录组测序公共数据GSE131907和人肺癌实体瘤单细胞转录组测序公共数据GSE203360,对数据进行聚类、差异基因本体功能富集分析,明确应用MPE建模的可行性。同时收集肺癌患者的MPE样本,经离心、裂解红细胞等富集细胞操作后,将其植入非肥胖型糖尿病重症联合免疫缺陷(non-obese diabetic/severe combined immunodeficient,NOD/SCID)小鼠皮下,待移植瘤生长至1000 mm³时进行瘤体传代及保存。对稳定传代移植瘤进行组织病理学检测,通过苏木精-伊红染色(hematoxylin-eosin staining,H-E染色)观察细胞组织形态,免疫组织化学法(immunohistochemistry,IHC)检测肺癌标志物表达情况。结果·经单细胞数据分析发现MPE中肿瘤细胞的增殖功能更强,提示MPE中肿瘤细胞PDX建模或具备更佳成瘤效果;共收集35例肺癌MPE样本,成功构建13例PDX模型,成功率达37.14%;在组织病理学检测中,H-E染色可见移植瘤组织细胞异型性明显,IHC检测显示细胞角蛋白7(cytokeratin 7,CK7)、甲状腺转录因子1(thyroid transcription factor-1,TTF1)和天冬氨酸蛋白酶A(Napsin A)等肺癌标志物均呈阳性表达。结论·通过富集肺癌患者MPE中的肿瘤细胞,成功构建了更为简便高效、可实时动态建模的PDX模型。该模型保留了肺癌患者肿瘤细胞的恶性特征及蛋白表达特性,为肺癌伴MPE患者的基础研究和临床用药指导提供了重要的实验模型工具。

转导蛋白β样1X连接受体1表达对卵巢癌A2780细胞增殖和迁移的影响

目的:研究转导蛋白β样1X连接受体1(transducin beta-like 1X-linked receptor,TBL1XR1)在卵巢癌患者组织中表达,及其对卵巢癌A2780细胞增殖和迁移的影响。方法:采用实时荧光定量PCR(qRT-PCR)检测10对卵巢癌组织、癌旁组织中及人卵巢癌IOSE80、A2780、CP70、SKOV-3中TBL1XR1 mRNA表达,筛选TBL1XR1 mRNA高表达细胞株。选择4~6周龄雌性BALB/C裸鼠,建立卵巢癌人源肿瘤异种移植(patient-derived tumor xenografts,PDTX)模型;将10只模型鼠均分为siR-NC组和si-TBL1XR1组,每组5只,分别给予siR-NC、si-TBL1XR1局部注射,10 mg/kg,每3 d注射1次,18 d后取各组瘤组织,计算其体积与重量。取卵巢癌A2780细胞,将其分为siR-NC组、si-TBL1XR1组、pcDNA3.1组和pcDNA3.1-TBL1XR1组,分别予以siR-NC、si-TBL1XR1、pcDNA3.1空载质粒和pcDNA3.1-TBL1XR1质粒处理;采用蛋白免疫印迹法检测各组卵巢癌细胞周期蛋白表达,MTT比色法检测细胞活力,流式细胞术检测细胞周期和凋亡细胞比例,以及Transwell细胞迁移实验检测细胞迁移能力。结果:卵巢癌组织中TBL1XR1 mRNA表达明显高于癌旁组织(P<0.05);人卵巢癌A2780细胞系TBL1XR1 mRNA表达明显高于卵巢癌IOSE80、CP70、SKOV-3细胞系(P<0.05)。与siR-NC组相比,第18天si-TBL1XR1组瘤体积明显减小(P<0.05),重量明显降低(P<0.05)。与siR-NC组相比,si-TBL1XR1组促癌细胞周期蛋白表达明显降低(P<0.05),与pcDNA3.1组相比,pcDNA3.1-TBL1XR1组表达则明显升高(P<0.05);与siR-NC组相比,si-TBL1XR1组卵巢癌细胞迁移数明显降低(P<0.05),早期凋亡和晚期凋亡细胞比例明显升高(P<0.05);与pcDNA3.1组相比,pcDNA3.1-TBL1XR1组卵巢癌细胞迁移数明显增多(P<0.05),早期凋亡和晚期凋亡细胞比例明显降低(P<0.05)。结论:TBL1XR1在卵巢癌组织中呈高表达,降低TBL1XR1 mRNA表达可抑制卵巢癌A2780细胞增殖和迁移。

基于患者源性口腔鳞状细胞癌类器官的药敏分析与初步临床应用

目的:基于所建立的患者源性口腔鳞状细胞癌类器官模型进行药物敏感性实验,对相应患者制定个性化药物治疗方案,观察其疗效与实验结果的匹配性。方法:本研究将通过活检或根治手术获得新鲜口腔鳞状细胞癌标本建立患者源性类器官模型,依托此模型对6种治疗口腔鳞状细胞癌药物(顺铂、紫杉醇、5-FU、西妥昔单抗、阿培利司、Nutlin-3)的敏感性进行测定,并对其中4例活检患者参考实验结果制定个性化用药方案,观察两个疗程后的近期治疗效果。结果:本研究成功建立了10例OSCC类器官(成功率10/12,83.3%),均可稳定传代、扩增达4代以上,且与亲本肿瘤组织表现出高度一致的组织病理学特征。口腔鳞状细胞癌类器官模型在药敏实验结果中表现出个体化差异,4例接受药物治疗的患者肿瘤控制结果均达到部分缓解标准,与实验结果一致。结论:本实验建立的患者源性口腔鳞状细胞癌类器官模型高度还原了同源亲本肿瘤的组织病理学特点,基于该模型的体外药敏实验与相应患者临床治疗反应一致,为建立个性化口腔鳞状细胞癌精准药物治疗新体系奠定基础。

转移性结直肠癌类器官的应用与展望

结直肠癌(colorectal cancer,CRC)是当前最常见的恶性肿瘤之一,具有较高的发病率和死亡率。50%以上的患者最终会出现肿瘤转移,且转移性结直肠癌(metastatic colorectal cancer,mCRC)患者对大部分药物敏感性不高,导致患者预后较差。虽然利用细胞系和患者源性异种移植模型进行mCRC的研究已较为成熟,但两者仍有较大的局限性。而患者来源肿瘤类器官作为患者衍生的三维培养物,具有模拟肿瘤体内特征、保留肿瘤细胞异质性、培养周期短及易于保存等优势,在mCRC的基础研究、药物作用机制的阐释、新药开发和个体化治疗等方面均展示出了极大的应用价值。本文概述了近年类器官技术在mCRC领域中的研究和挑战,旨在阐明类器官在mCRC中的研究价值,为后续研究提供方向。

人源胰腺癌类器官模型的构建及应用新进展

胰腺癌严重威胁人类健康,其高度异质性和恶性表型使患者的5年生存率仅为7.2%。胰腺癌的发病机制及药物开发研究常依赖于传统的二维培养模型(细胞系)和患者来源异种移植瘤模型,但因细胞系缺乏肿瘤的三维环境和异质性特征,而异种移植瘤模型则存在培养时间长、成功率低、难以开展高通量药物筛选等问题,急需开发可高度反映胰腺癌特征和分子变异的三维培养模型。人源胰腺癌类器官作为近年发展起来的三维培养模型,是从组织样本中提取的多细胞单位,在机械和酶消化后嵌入细胞外基质凝胶中,可再现原患者的组织学特征及器官特点,甚至具有原器官的功能。随着胰腺癌类器官培养体系的不断发展及完善,其简便、经济及稳定的培养技术已经逐步建立,推动人源胰腺癌类器官的应用扩展到药物高通量筛选、个体化精准治疗、更深入的发病机制及针对性的药物开发研究。同时,人源胰腺癌类器官亦是胰腺癌临床分子分型研究的优势全新半体内模型,特别在针对研究高突变及高异质性患者的病因、分子特征、组织形态及体细胞突变负荷方面表现优越。大样本量人源胰腺癌类器官生物样本库的构建,将成为生物学、基础医学和临床肿瘤学研究的全新平台,有助于胰腺癌发病机制的深入研究及治疗策略的优化。人源胰腺癌类器官可再现临床患者特征的实验室模型的应用,将使药物筛选、发病机制研究、个体化治疗的实现成为可能。本文综述人源胰腺癌类器官的最新研究进展,希望该文为从事相关人源胰腺癌类器官研究的人员提供借鉴。

双硫死亡相关基因对结直肠癌预后和药物敏感性的预测价值

目的通过生物信息学方法建立基于双硫死亡相关基因表达量预测结直肠癌(colorectal cancer,CRC)预后和药物敏感性的评分模型,并结合CRC患者来源类器官(CRC patient-derived organoids,CRC-PDOs)验证。方法利用NMF(non-negative Matrix Factorization)算法、Cox和LASSO回归分析明确对CRC预后有预测价值的双硫死亡相关基因,构建双硫死亡相关风险评分公式,结合GO(Gene Ontology)和KEGG(Kyoto Encyclopedia of Genes and Genomes)分析不同集群之间的差异基因及富集通路。通过GDSC数据库预测不同分组CRC患者对化疗药物的敏感性,并利用CRC-PDOs进行验证。结果NMF算法结果显示基于双硫死亡相关基因可建立CRC患者分群,COX回归分析显示LRPPRC和SLC7A11是能够预测CRC患者预后的潜在基因(P=0.047、0.033),CRC患者肿瘤中的SLC7A11低表达或LRPPRC高表达与总生存率(overall survival,OS)显著相关(P=0.004、0.003)。基于LASSO回归分析,双硫死亡风险评分公式为:Risk score=LRPPRC×(-0.6705)+SLC7A11×0.3112,且能将GSE161158数据集分群为高风险和低风险组。双硫死亡高低危群集间有125个差异表达基因(differentially expressed genes,DEGs),结合GO和KEGG结果,高危群集患者的上调基因主要富集于免疫调节,如白细胞趋化性、粒细胞迁移和Toll样受体结合。低风险集群的特征是与硫化物代谢和运输相关的途径,如硫化物跨膜转运蛋白活性。基于GDSC数据库,双硫死亡相关的双基因表达水平可预测化疗药物敏感性,化疗药物(伊立替康)对于CRC-PDOs的生长抑制结果提示患者的结直肠癌组织中SLC7A11和LRPPRC的相对表达量与其类器官映射疗效呈线性相关(P=0.007、0.040)。结论根据生物信息学分析和CRC-PDOs实验结果,双硫死亡相关评分能够预测结直肠癌患者预后和药物敏感性,具有潜在的临床应用前景。

Patient-derived xenograft platform of OSCC： a renewable human bio-bank for preclinical cancer research and a new co-clinical model for treatment optimization

Advances in next-generation sequencing and bioinformatics have begun to reveal the complex genetic landscape in human cancer genomes, including oral squamous cell carcinoma （OSCC）. Sophisticated preclinical models that fully represent intra- and inter-tumoral heterogeneity are required to understand the molecular diversity of cancer and achieve the goal of personalized therapies. Patient-derived xenograft （PDX） models generated from human tumor samples that can retain the histological and genetic features of their donor tumors have been shown to be the preferred preclinical tool in translational cancer research compared with other conventional preclinical models. Specifically, genetically well-defined PDX models can be applied to accelerate targeted antitumor drug development and biomarker discovery. Recently, we have successfully established and characterized an OSCC PDX panel as part of our tumor bio-bank for translational cancer research. In this paper, we discuss the establishment, characterization, and preclinical applications of the PDX models. In particular, we focus on the classification and applications of the PDX models based on validated annotations, including clinicopathological features, genomic profiles, and pharmacological testing information. We also explore the translational value of this well-annotated PDX panel in the development of co-clinical trials for patient stratification and treatment optimization in the near future. Although various limitations still exist, this preclinical approach should be further tested and improved.

解毒疏肝汤联合隔姜灸对老年肝癌介入治疗患者的影响

目的 探究解毒疏肝汤联合隔姜灸对老年肝癌介入治疗患者的近期疗效及对血管内皮生长因子(VEGF)、白介素-8(IL-8)、血小板衍生生长因子(PDGF)的影响。方法 选取2020年2月至2023年2月期间在本院进行介入治疗的74例原发性肝郁脾虚型肝癌患者,以随机数表法将其均分为两组,对照组(常规水化、保肝、止吐)及观察组(在常规水化、保肝、止吐的基础上应用解毒疏肝汤联合隔姜灸)各37例,对比两组临床症状(发热、呕吐、肝区疼痛、腹胀、黄疸)改善时间、中医证候积分、肝功能[丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、直接胆红素(DBIL)、总胆红素(TBIL)]、炎症因子[白细胞介素-8(IL-8)、白细胞介素-1(IL-1)、肿瘤坏死因子(TNF-α)]、血管内皮生长因子(VEGF)、血小板衍生生长因子(PDGF)、碱性成纤维细胞生长因子(bFGF)。结果 观察组发热、呕吐、肝区疼痛、腹胀、黄疸改善时间短于对照组,差异有统计学意义(P <0.05)。治疗后,观察组主证、次症、总分低于对照组,差异有统计学意义(P <0.05)。治疗后,观察组ALT、AST、DBIL、TBIL水平低于对照组,差异有统计学意义(P <0.05)。治疗后,观察组IL-8、IL-1、TNF-α水平低于对照组,差异有统计学意义(P <0.05)。治疗后,观察组VEGF、PDGF、bFGF水平低于对照组,差异有统计学意义(P <0.05)。结论 在常规水化、保肝、止吐的基础上实施解毒疏肝汤联合隔姜灸对老年肝癌介入治疗效果显著,可改善患者肝功能、血管内皮功能,降低炎症因子、VEGF、PDGF、bFGF水平。

人源肿瘤异种移植小鼠模型研究进展

建立适当的移植瘤模型对于癌症研究至关重要。迄今为止,最常用的移植瘤模型是人源肿瘤细胞系异种移植模型(cancer cell line-based xenograft,CDX),即将体外传代培养的肿瘤细胞移植到免疫缺陷小鼠体内形成移植瘤。虽然这种模型容易建立且建模周期较短,但不能充分代表临床癌症患者,因此,具有能够稳定保留肿瘤异质性的人源肿瘤异种移植模型(patient-derived tumor xenograft,PDX)在诸多应用中开始代替CDX模型。PDX模型通过将患者组织或原代细胞直接植入免疫缺陷小鼠进行成瘤,这种方式保留亲代肿瘤的组织病理学、分子特征和药物反应性,可作为临床前模型,在药物筛选、生物标志物开发和联合临床试验等方面具有显著优势。在这篇综述中,我们将详细介绍PDX模型的构建方法和应用,总结模型建立过程中及进行临床前研究中可能遇到的问题。

结直肠癌PDX模型成瘤率的影响因素及临床意义

目的建立结直肠癌(CRC)患者来源的肿瘤组织异种移植(PDX)模型,评估PDX模型成瘤率的影响因素,并初步进行化学治疗实验。方法选取2019年11月至2020年10月绍兴市人民医院择期手术CRC患者。将手术获取的肿瘤组织接种于NSG小鼠右侧腰背部,建立PDX模型,并传至F3代,分析PDX模型成瘤率的影响因素;化学治疗药物选择5-氟尿嘧啶、奥沙利铂以及丙泊酚。结果本研究共纳入60例CRC患者,PDX模型成瘤为37例,成瘤率62%;平均成瘤时间为(34±12)d;原发瘤恶性程度(CRC分期和细胞分化程度)、术前癌胚抗原(CEA)水平以及肿瘤位置等因素影响PDX模型成瘤率(P<0.01)。CRC-PDX移植瘤组织与患者肿瘤组织生物学特征高度一致。4种化学治疗方案均能抑制肿瘤生长,致肿瘤组织破坏,丙泊酚可以抑制小鼠腹泻,对肠黏膜具有保护作用。结论本研究建立的CRC-PDX模型,较好地保持原发肿瘤的生物学特性,可作为CRC患者个体化治疗的参考模型。原发肿瘤恶性程度是PDX模型成瘤率的主要影响因素。

人胶质母细胞瘤细胞系Y1203的构建及应用

目的构建一株具备干细胞表型的胶质母细胞瘤细胞系Y1203,用于研究肿瘤细胞干性维持以及治疗耐药的潜在机制。方法Y1203细胞来源于一位49岁女性胶质母细胞瘤患者复发再手术切除的肿瘤组织。使用Y1203细胞构建人源胶质母细胞瘤原位及皮下移植(patient derived tumor xenograft,PDX)模型。采用流式细胞术检测不同代数Y1203细胞的细胞周期。采用实时荧光定量聚合酶链反应检测Y1203细胞的干细胞标志物转录因子SOX-2(transcription factor SOX-2,SOX2)和细胞黏附分子分化抗原簇-44(cluster of differentiation-44,CD44)表达。基于转录组测序技术(RNA sequencing,RNA-Seq)比较Y1203细胞和U87细胞表达谱差异。结果短串联重复序列(short tandem repeats,STR)鉴定确认Y1203细胞与患者肿瘤组织同源,且其与ExPASy数据库中涵盖的常见细胞系完全不同。基因检测结果提示异柠檬酸脱氢酶1(isocitrate dehydrogenase 1,IDH1)无突变,端粒酶反转录酶(telomerase reverse transcriptase,TERT)启动子突变,原癌基因(B-Raf proto-oncogene,BRAF V600E)、抑癌基因(tumor protein p53,TP53)和转录调控因子(ATRX chromatin remodeler,ATRX)基因突变。人胶质母细胞瘤细胞系Y1203可实现在体外无限增殖并用于建立PDX模型。与其他胶质瘤细胞系相比,Y1203细胞中高表达的肿瘤干细胞标志物为SOX2和CD44,并显著富集胶质瘤干细胞相关通路。结论人胶质母细胞瘤细胞系Y1203分化低,恶性程度较高,对放射性治疗和化学治疗抵抗性强,具有胶质母细胞瘤干细胞特性。该细胞系的建立为胶质母细胞瘤临床前研究提供了实验基础。