小鼠Lewis肺癌原位模型的构建

背景与目的肺癌原位模型包括小鼠自发性肺肿瘤模型和气道内接种模型等,但自发性肺肿瘤模型耗时较长且成瘤率不能保证,而气道内接种模型成瘤部位及大小不稳定。本研究以3LL细胞系细胞接种于C57BL/6小鼠肺原位,与皮下接种模型比较,探讨其稳定性、转移特性,并建立小鼠肺原位癌模型的优化方法。方法将不同数量级的3LL细胞分别直接接种于C57BL/6小鼠左侧腋下制备皮下接种模型和以Matrigel悬浮后接种于其左肺制备原位接种模型,观察两种模型的生存期,并对小鼠进行解剖后行病理切片检查、免疫组化染色检测微血管密度、流式细胞仪检测CD44v。结果皮下组成瘤率分别为100%、66.7%、16.7%,未见明显转移。原位组成瘤率分别为100%、100%、83.3%,并可转移至对侧胸廓及肺脏。原位组中位生存期(38d、35d、23d)明显少于皮下接种组(82d、72d、50d)。原位接种组微血管密度(120.2±9.73)高于皮下组(92.6±7.12)。原位接种组肿瘤细胞悬液CD44v表达(26.46±1.56)%高于皮下接种组(23.13±1.02)%。结论以3LL细胞接种于小鼠肺部所建立的肺癌原位模型简单可靠,重复性高,具有较...     

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。     

人ACHN肾癌裸鼠移植瘤模型的建立及其特性研究

目的　用ACHN肾癌细胞株,建立人肾癌裸小鼠异位移植模型。方法　将ACHN人肾癌细胞株2. 0×10 /0. 2ml接种于裸小鼠背部皮下,观察移植肿瘤的生长状况、移植成功率、肺转移率及移植瘤病理形态,检测肿瘤细胞的增殖与凋亡。结果　异位移植成功率为 75%,肺转移率 58. 3%,肿瘤细胞增殖指数(PI)为 54. 84±11. 39,凋亡指数(AI)为 1. 75±0. 83,AI/PI与肿瘤体积呈负相关 (r=-0. 8332,P<0. 05)。皮下瘤病理形态与肾腺癌相似。结论　人ACHN肾癌裸鼠异位移植瘤模型可作为肾癌及其转移的研究工具。     

小鼠Lewis肺癌胸腔与皮下移植模型的比较研究

目的:建立小鼠Lewis肺癌胸腔移植模型与皮下移植模型,比较两种模型生存时间、生活状态以及肿瘤进展情况,寻找适合评价肺癌发展进程的模型。方法:将传代培养的Lewis肺癌细胞(LLC)接种于C57BL/6近交系小鼠的腋后线第6肋间胸腔内和腋后部皮下,建立两组肺癌模型:胸腔组和皮下组。观察两组模型的成瘤率、小鼠的存活时间和生活状态及肿瘤的浸润和转移情况。结果:小鼠Lewis肺癌胸腔组和皮下组接种成瘤率100%。胸腔组小鼠中位生存时间13.5(8～18)d,皮下组中位生存时间45(25～71)d,胸腔组的生存期范围离散程度较皮下组小,P=0.023。胸腔组小鼠在生存过程中可出现肿瘤局部浸润和远处转移的晚期肺癌特征,而皮下组则很少出现上述情况。病程中胸腔组小鼠恶病质状态较皮下组严重,胸腔组小鼠体质量下降、饲料消耗量、游泳试验和常压耐缺氧试验时间均较皮下组严重,差异有统计学意义。结论:小鼠Lewis肺癌胸腔移植模型较皮下移植模型更能反映肺癌发展的进程,胸腔移植模型可作为晚期肺癌的动物模型。     

小鼠Lewis肺癌胸腔与皮下移植模型的比较研究

目的：建立小鼠Lewis肺癌胸腔移植模型与皮下移植模型，比较两种模型生存时间、生活状态以及肿瘤进展情况，寻找适合评价肺癌发展进程的模型。方法：将传代培养的Lewis肺癌细胞（LLC）接种于C57BL／6近交系小鼠的腋后线第6肋间胸腔内和腋后部皮下，建立两组肺癌模型：胸腔组和皮下组。观察两组模型的成瘤率、小鼠的存活时间和生活状态及肿瘤的浸润和转移情况。结果：小鼠Lewis肺癌胸腔组和皮下组接种成瘤率100％。胸腔组小鼠中位生存时间13．5（8～18）d，皮下组中位生存时间45（25～71）d，胸腔组的生存期范围离散程度较皮下组小，P=0．023。胸腔组小鼠在生存过程中可出现肿瘤局部浸润和远处转移的晚期肺癌特征，而皮下组则很少出现上述情况。病程中胸腔组小鼠恶病质状态较皮下组严重，胸腔组小鼠体质量下降、饲料消耗量、游泳试验和常压耐缺氧试验时间均较皮下组严重，差异有统计学意义。结论：小鼠Lewis肺癌胸腔移植模型较皮下移植模型更能反映肺癌发展的进程，胸腔移植模型可作为晚期肺癌的动物模型。     

BNX小鼠人胃癌原位移植模型的建立及其生物学特性的观察

目的应用T、B、NK细胞联合免疫缺陷的BNX小鼠建立人胃癌原位移植高转移模型.方法将MKN-45细胞株接种至BNX小鼠皮下,成瘤后将肿瘤组织剪成小块通过手术移植至BNX小鼠的胃壁,观察肿瘤生长与转移情况,8周后对动物进行解剖.结果胃壁原位移植瘤浸润破坏胃壁各层组织结构,并直接扩散至周围脏器.肿瘤的移植成功率为100%(23/23),表现出很高的侵袭和转移特性:原位肿瘤平均体积2837.07±1044.04 mm3,局部和远处淋巴结转移率达83%(19/23),肝转移率为83%(19/23),肺转移率为65%(15/23),膈转移率为39%(9/23),发生腹腔种植与腹水的动物为35%(8/23),此外,还有个别脾转移.结论通过T、B、NK细胞联合免疫缺陷的BNX小鼠建立人胃癌原位移植模型可以更好地模拟人胃癌侵袭与转移本身的自然过程,对人胃癌防治及其转移机理的研究提供一个更为理想的模型.     

人骨肉瘤原位移植模型的建立及生物学特征

目的 用人骨肉瘤细胞系HOS-98建立人骨肉瘤裸鼠胫骨原位移植模型,以探讨宿主器官微环境对人骨肉瘤细胞侵袭及转移等生物学行为的影响。方法 将人骨肉瘤细胞系HOS-98接种于裸鼠皮下,形成移植瘤,用传代移植瘤组织作为移植材料,进行胫骨原位移植及皮下移植。分别于移植后4周和8周处死小鼠,进行病理形态学检查,并对两种方法在成瘤率、生长方式及侵袭、转移等生物学行为比较。结果 两种移植方式在成瘤率及形态学上无明显不同,胫骨原位移植的潜伏期较短,并且生长快于皮下移植方式。皮下移植瘤呈局限性膨胀生长,有不完整的纤维包膜,瘤内类骨基质较少见,未见肺转移,观察8周时无明显消瘦;而胫骨原位移植瘤侵袭周围组织,可见发生肺转移,8周明显消瘦。原位移植的裸鼠血清ALP水平高于皮下移植者。原位移植的X线检查有明显的类似于人的骨性反应。结论 用人骨肿瘤细胞系HOS-98皮下接种的移植瘤作为移植材料是建立肿瘤异位移植的可行途径,裸鼠胫骨微环境较皮下组织更适合于人骨肉瘤的侵袭及转移表达,裸鼠胫骨原位移植模型的恶性生物学行为更接近临床骨肉瘤患者的体内侵袭及转移实际,该原位移植模型为今后的实验研究提供了更加接近患者实际的实验模型。     

EGFP标记的人肺癌裸鼠原位移植模型的建立

背景与目的 小鼠活体分子成像模型可以连续实时监测活体肿瘤的变化.本研究拟通过外科原位移植法建立表达绿色荧光蛋白的肺癌裸鼠原位移植模型并探讨其肿瘤生物学特性,从而建立一个良好的肺癌动物实验研究平台.方法 利用逆转录病毒转染法将增强型绿色荧光蛋白基因导人人肺癌大细胞系NCI-H460,采用外科原位移植法建立肺癌原位移植模型.定期通过小动物活体荧光成像系统观察肿瘤生长,利用相关性检验分析荧光面积和肿瘤体积之间的相关关系,并观察原位移植术后裸鼠的生存期和肿瘤转移情况.结果 模型建立后1周通过皮瓣在荧光体视镜下可观察到肺部肿瘤的绿色荧光,成瘤率为100%.荷瘤裸小鼠平均生存期为34.2天.解剖裸鼠观察到肿瘤侵及对侧肺、纵隔及肺门淋巴结、胸膜和膈肌,转移率分别为87.596、75%、25%和12.5%.肿瘤体积和荧光面积具有相关性(r=0.873,P=0.001).结论 外科原位移植法建立的表达EGFP的裸鼠肺癌原位模型是肺癌临床前研究的理想的实验工具.应用小动物活体荧光成像系统能够定量客观评价肿瘤在动物体内的生长、侵袭和转移,该模型可应用于肺癌的基础研究和新药开发.     

羟基红花黄色素A对小鼠Lewis肺癌移植瘤的抑制作用

目的 研究羟基红花黄色素A(hydroxysafflor yellow A,HSYA)对C57BL/6小鼠Lewis肺癌移植瘤的血管抑制作用.方法 C57BL/6近交系小鼠30只,造模后,分为5组,每组6只,分别使用生理盐水、环磷酰胺(CTX)及HSYA(分低、中、高3个剂量)作用于动物模型.观察各组肿瘤体积大小,分析小鼠生长曲线,并在造模第22天处死所有小鼠,无菌条件下剥取瘤组织进行HE染色,观察不同浓度HSYA在不同视野下对肿瘤血管的抑制情况.结果 HSYA中、小剂量组对小鼠移植瘤血管生成有一定抑制作用.结论 中药羟基红花黄色素A(HSYA)对Lewis肺癌小鼠移植瘤血管生成有一定的抑制作用.     

人肺癌高转移动物模型的筛选及其细胞系的建立

目的:建立人肺癌小鼠高转移模型及高转移细胞系,同时观察相关生物学特性,为肺癌转移机制和防治等研究提供有用的实验工具。方法:切除首代小鼠移植瘤,以延长动物生存时间而获得转移灶,从第二代起采用肺转移灶→皮下移植→肺转移灶→皮下移植的体内循环筛选方法建立NOD/SCID小鼠人肺癌细胞SPC-A-1皮下移植瘤高转移模型,并进行肿瘤的生长和转移情况、组织病理学观察,同时建立相应的高转移细胞系,进行各种相关生物学特性观察。结果:第一代移植瘤切除后转移率达66.7%,通过4代体内反复筛选建立了100%肺转移NOD/SCID小鼠模型及相应高转移细胞系,细胞生长行为和染色体分析等生物学特性观察表明该细胞系保持了原有的人肺腺癌的生物学特性。结论:应用体内筛选的方法成功建立了人肺癌皮下移植瘤高转移模型及高转移细胞系,为肺癌防治研究及抗转移实验治疗提供了理想的动物模型。     

Mouse colostomy model for studies on large bowel cancer

The development of appropriate animal model systems has been proposed as a means of facilitating the study of human colorectal cancer. This report describes the development and use of a blind-colorectal pouch in a carcinoma mouse model. The blind-pouch was prepared in C57BL/6J mice by surgically exteriorizing the descending colon and producing two stomata in the abdominal wall. The proximal stoma served as an end colostomy and the distal stoma created as a mucous fistula. Surgical closure of the anus thus provided a colorectal pouch. In pilot studies it was found that N-methyl-N-nitrosourea (MNU) and radiation together but neither separately produced tumors in the pouch of surviving mice. Further, inoculation of C38 syngeneic tumor cell suspension into the pouch and immediate closure resulted in tumor takes within three to four weeks. The use of this model in carcinogenesis and the immunology of colon cancer simulate the human colorectal cancer problem more closely than previous animal models.     

Nude mouse model to study passive humoral immunotherapy directed against B16 F10 murine melanoma

A model to study passive humoral immunotherapy of experimental melanoma was generated by subcutaneous injection of B16 F10 murine melanoma cells in the midtail of BALB/C nude (nu/nu) mice. Mice were challenged with melanoma cells pretreated: (1) with complete culture medium, (2) with 10% adjuvant control serum, (3) with 10% anti-fECA (formalinized extracellular antigens) immune serum, or (4) with a monoclonal antibody (mAb H2-3-3) specific for the B700 melanoma-associated antigen. All control mice challenged with melanoma cells pretreated either with culture medium or with medium containing adjuvant control serum (Groups I and II) died during the observation period of 84 days. At day 84, 60% of the mice challenged with melanoma cells pretreated with anti-fECA immune serum (Group III) survived, as did 100% of the mice challenged with cells pretreated with mAb H2-3-3 (Group IV). Injection of melanoma cells pretreated with mAb H2-3-3 was associated with the greatest reduction of subsequent local tumor growth and the lowest number of metastatic lung tumors. The inhibitory effects of immune sera in vivo also correlated with in vitro effects of anti-fECA immune serum and mAb H2-3-3, determined on B16 F10 melanoma target cells using assays for DNA synthesis and antibody dependant cellular cytotoxicity (ADCC). In sum, this nude mouse model for the study of passive humoral immunotherapy of experimental melanoma was utilized to demonstrate significant protective effects against B16 F10 melanoma cell challenge by treatment with anti-fECA immune sera or a melanoma-specific monoclonal antibody. © 1994 Wiley-Liss, Inc.     

Optimization of an orthotopic murine model of head and neck squamous cell carcinoma in fully immunocompetent mice--role of toll-like-receptor 4 expressed on host cells

For preclinical studies of immune-modulating anticancer drugs a murine model that attempts to parallel the clinical nature of head and neck cancer in fully immunocompetent mice is required. In this study we compared features of the squamous cell carcinoma (SCC) VII model after subcutaneous (back, flank) and orthotopic (floor of mouth) injection both in fully immunocompetent C3H/HeN and in previously studied C3H/HeJ mice, which harbor a functional toll-like receptor 4 (TLR-4) deficiency. As C3H/HeN mice do not harbor this deficiency, the presented murine model is an optimization of previously described C3H/HeJ models, which, because of the TLR-4-deficiency, have inherent drawbacks for tumor immunologic studies. We found that tumor growth was accelerated and tumor incidence was increased by about 20% after s.c. injection in TLR-4-deficient mice. Strikingly, tumor-related weight loss (cachexia) was more pronounced in fully immunocompetent C3H/HeN mice (26%) versus TLR-4-deficient C3H/HeJ mice (7.9% weight loss) at high tumor dose. Orthotopic tumors were biologically distinct from subcutaneous tumors as they showed accelerated growth and a distinct immune cell infiltrate. We conclude that a model of orthotopic implantation of SCC VII tumor cells into fully immunocompetent syngeneic C3H/HeN mice reflects features of human head and neck cancer and provides a valuable experimental model for immunological studies in this tumor entity. Our data suggest that TLR-4 expressed by host cells is involved in the regulation of tumor-related cachexia and tumor control.     

Characteristics of Paecilomyces lilacinus infection comparing immunocompetent with immunosuppressed murine model

The characteristics of Paecilomyces lilacinus infection were evaluated using two murine experimental models: immunocompetent and immunosuppressed. The evaluation criteria for characteristics of infection were clinical signs, weight loss, survival rates, histopathological alterations and the number of viable fungal cells re-isolated from different organs; and those for immunological status were in vitro lymphoproliferative response, cell surface phenotyping and IFN-γ production. Morphological evaluation showed that P. lilacinus isolates presented morphological characteristics consistent with those described in the literature. The immunocompetent mice could be infected by the fungi, but they did not develop the disease, unlike the immunosuppressed mice, which showed clinical signs of mycosis in an environment of suppressed cellular immune response. The hypothesis of latent infection reactivation in mice was not confirmed. The difference observed in the infection rate of the two fungi isolates points to an intrinsic variation between strains of P. lilacinus and led us to hypothesise that even in the presence of immunosuppressed environment, the fungus virulence can play a role in the pathogenesis of hyalohyphomycosis.     

C-kit signaling promotes proliferation and invasion of colorectal mucinous adenocarcinoma in a murine model

It was reported that the receptor tyrosine kinase (RTK) family often highly expressed in several mucinous carcinomas. In the present study, we established a murine model of colorectal mucinous adenocardinoma (CRMAC) by treating C57 mice [both wild type (WT) and loss-of-function c-kit mutant type (Wads^−/−)] with AOM+DSS for 37 weeks and found that c-kit, a member of RTK family, clearly enhanced the tumor cell proliferation by decreasing p53 and increasing cyclin D1 through AKT pathway. Significantly, c-kit strongly promoted tumor cell invasiveness by increasing ETV4, which induced MMP7 expression and epithelial-mesenchymal transition (EMT) via ERK pathway. In vitro up- or down-regulating c-kit activation in human colorectal cancer HCT-116 cells further consolidated these results. In conclusion, our data suggested that the c-kit signaling obviously promoted proliferation and invasion of CRMAC. Therefore, targeting the c-kit signaling and its downstream molecules might provide the potential strategies for treatment of patients suffering from CRMAC in the future.     

Ineffectiveness of Doxorubicin Treatment on Solitary Dormant Mammary Carcinoma Cells or Late-developing Metastases

Breast cancer is noted for long periods of tumor dormancy and metastases can occur many years after treatment. Adjuvant chemotherapy is used to prevent metastatic recurrence but is not always successful. As a model for studying mechanisms of dormancy, we have used two murine mammary carcinoma cell lines: D2.0R/R cells, which are poorly metastatic but form metastases in some mice after long latency times, and D2A1/R cells, which form more numerous metastases much earlier. Previously we identified a surprisingly large population of dormant but viable solitary cells, which persisted in an undivided state for up to 11 weeks after injection of D2.0R/R cells. Dormant cells were also detected for D2A1/R cells, in a background of growing metastases. Here we used this model to test the hypothesis that dormant tumor cells would not be killed by cytotoxic chemotherapy that targets actively dividing cells, and that the late development of metastases from D2.0R/R cells would not be inhibited by chemotherapy that effectively inhibited D2A1/R metastases. We injected mice with D2A1/R or D2.0R/R cells via a mesenteric vein to target liver. We developed a doxorubicin (DXR) treatment protocol that effectively reduced the metastatic tumor burden from D2A1/R cells at 3 weeks. However, this treatment did not reduce the numbers of solitary dormant cells in mice injected with either D2A1/R or D2.0R/R cells. Furthermore, DXR did not reduce the metastatic tumor burden after an 11-week latency period in mice injected with D2.0R/R cells. Thus, apparently effective chemotherapy may spare non-dividing cancer cells, and these cells may give rise to metastases at a later date. This study has important clinical implications for patients being treated with cytotoxic chemotherapy.     

Animal models of osteosarcoma

Appendicular osteosarcoma (OS) is a primary mesenchymal tumor arising from malignantly transformed osteoblasts. In people, OS is the most common nonhematopoietic, primary skeletal neoplasm diagnosed in adolescents and is the second leading cause of cancer-related fatalities within this age group. Despite aggressive therapeutic management, including limb-sparing surgeries and dose-intense systemic chemotherapies, 30–40% of patients will experience progressive metastatic disease within 5 years of diagnosis. In order to reduce the fatality rate associated with recurrent or metastatic OS, a more thorough understanding of OS pathogenesis and biology is required. Towards this pursuit, comparative animal models of OS have been developed and are actively being studied to expand our fundamental understanding of OS. It is anticipated that specific animal models of OS, which most accurately recapitulate the natural disease process in people, will be most useful for advancing our understanding of OS biology, and will facilitate the discovery of disease pathogenesis and the identification of novel therapeutic strategies for managing this lethal metastatic bone sarcoma.     

A novel systemically administered toll‐like receptor 7 agonist potentiates the effect of ionizing radiation in murine solid tumor models

Although topical TLR7 therapies such as imiquimod have proved successful in the treatment of dermatological malignancy, systemic delivery may be required for optimal immunotherapy of nondermatological tumors. We report that intravenous delivery of the novel small molecule TLR7 agonist, DSR‐6434, leads to the induction of type 1 interferon and activation of T and B lymphocytes, NK and NKT cells. Our data demonstrate that systemic administration of DSR‐6434 enhances the efficacy of ionizing radiation (IR) and leads to improved survival in mice bearing either CT26 or KHT tumors. Of the CT26 tumor‐bearing mice that received combined therapy, 55% experienced complete tumor resolution. Our data reveal that these long‐term surviving mice have a significantly greater frequency of tumor antigen specific CD8⁺ T cells when compared to age‐matched tumor‐naïve cells. To evaluate therapeutic effects on spontaneous metastases, we showed that combination of DSR‐6434 with local IR of the primary tumor significantly reduced metastatic burden in the lung, when compared to time‐matched cohorts treated with IR alone. The data demonstrate that systemic administration of the novel TLR7 agonist DSR‐6434 in combination with IR primes an antitumor CD8⁺ T‐cell response leading to improved survival in syngeneic models of colorectal carcinoma and fibrosarcoma. Importantly, efficacy extends to sites outside of the field of irradiation, reducing metastatic load. Clinical evaluation of systemic TLR7 therapy in combination with IR for the treatment of solid malignancy is warranted.