Development of the transgenic cyan fluorescent protein (CFP)‐expressing nude mouse for “Technicolor” cancer imaging

A major goal for in vivo biology is to develop models which can express multiple colors of fluorescent proteins in order to image many processes simultaneously in real time. Towards this goal, the cyan fluorescent protein (CFP) nude mouse was developed by crossing non‐transgenic nude mice with the transgenic CK/ECFP mouse in which the β‐actin promoter drives expression of CFP in almost all tissues. In crosses between nu/nu CFP male mice and nu/+ CFP female mice, approximately 50% of the embryos fluoresced blue. In the CFP nude mice, the pancreas and reproductive organs displayed the strongest fluorescent signals of all internal organs which vary in intensity. Orthotopic implantation of XPA‐1 human pancreatic cancer cells expressing red fluorescent protein (RFP); or green fluorescent protein (GFP) in the nucleus and RFP in the cytoplasm, was performed in female nude CFP mice. Color‐coded fluorescence imaging of these human pancreatic cancer cells implanted into the bright blue fluorescent pancreas of the CFP nude mouse afforded novel insight into the interaction of the pancreatic tumor and the normal pancreas, in particular the strong desmoplastic reaction of the tumor. The naturally enhanced blue fluorescence of the pancreas in the CFP mouse serves as an ideal background for color‐coded imaging of the interaction of implanted cancer cells and the host. The CFP nude mouse will provide unique understanding of the critical interplay between the cancer cells and their microenvironment. J. Cell. Biochem. 107: 328–334, 2009. © 2009 Wiley‐Liss, Inc.     

Tumor-secreted miR-214 induces regulatory T cells: a major link between immune evasion and tumor growth

An increased population of CD4⁺CD25^highFoxp3⁺ regulatory T cells (Tregs) in the tumor-associated microenvironment plays an important role in cancer immune evasion. However, the underlying mechanism remains unclear. Here we observed an increased secretion of miR-214 in various types of human cancers and mouse tumor models. Tumor-secreted miR-214 was sufficiently delivered into recipient T cells by microvesicles (MVs). In targeted mouse peripheral CD4⁺ T cells, tumor-derived miR-214 efficiently downregulated phosphatase and tensin homolog (PTEN) and promoted Treg expansion. The miR-214-induced Tregs secreted higher levels of IL-10 and promoted tumor growth in nude mice. Furthermore, in vivo studies indicated that Treg expansion mediated by cancer cell-secreted miR-214 resulted in enhanced immune suppression and tumor implantation/growth in mice. The MV delivery of anti-miR-214 antisense oligonucleotides (ASOs) into mice implanted with tumors blocked Treg expansion and tumor growth. Our study reveals a novel mechanism through which cancer cell actively manipulates immune response via promoting Treg expansion.     

人卵巢癌裸鼠移植实体瘤模型的建立

目的建立人卵巢癌裸鼠移植实体瘤模型。方法将1例人卵巢癌组织移植裸鼠,建立人卵巢癌裸鼠原代移植实体瘤模型的基础上,再将实体瘤皮下移植、实体瘤原位移植、实体瘤细胞移植裸鼠。观察裸鼠实体瘤生长和转移情况,称量其体重、子宫卵巢重、瘤重及瘤的大小,并作病理、电镜、染色体检查。结果成功地建立人卵巢癌裸鼠移植实体瘤模型,并已传至第18代,传代移植成功率100％,组织学和超微结构形态均证明该实体瘤保留了原人卵巢癌特征,有人卵巢癌染色体特征,并出现肝、脾转移。结论本研究建立人卵巢癌裸鼠移植实体瘤模型与人相似,通过18代传代和实验观察方法稳定可靠。为人卵巢癌的研究提供了理想的动物模型。     

Temozolomide and Pazopanib Combined with FOLFOX Regressed a Primary Colorectal Cancer in a Patient-derived Orthotopic Xenograft Mouse Model

Purpose: The goal of the present study was to determine the efficacy of temozolomide (TEM) and pazopanib (PAZ) combined with FOLFOX (oxaliplatin, leucovorin and 5-fluorouracil) on a colorectal cancer patient-derived orthotopic xenograft (PDOX) mouse model. Materials and Methods: A colorectal cancer tumor from a patient previously established in non-transgenic nude mice was implanted subcutaneously in transgenic green fluorescence protein (GFP)-expressing nude mice in order to label the tumor stromal cells with GFP. Then labeled tumors were orthotopically implanted into the cecum of nude mice. Mice were randomized into four groups: Group 1, untreated control; group 2, TEM + PAZ; group 3, FOLFOX; group 4, TEM + PAZ plus FOLFOX. Tumor width, length, and mouse body weight were measured weekly. The Fluor Vivo imaging System was used to image the GFP-lableled tumor stromal cells in vivo. H&E staining and immunohistochemical staining were used for histological analysis. Results: All three treatments inhibited tumor growth as compared to the untreated control group. The combination of TEM + PAZ + FOLFOX regressed tumor growth significantly more effectively than TEM + PAZ or FOLFOX. Only the combination of TEM + PAZ + FOLFOX group caused a decrease in body weight. PAZ suppressed lymph vessels density in the colorectal cancer PDOX mouse model suggesting inhibition of lymphangiogenesis. Conclusion: Our results suggest that the combination of TEM + PAZ + FOLFOX has clinical potential for colorectal cancer patient.     

小鼠Lewis肺癌原位模型的建立

目的利用Matrigel与Lewis制备细胞混悬液注射于小鼠左肺内,建立小鼠Lewis肺癌原位模型,评价其肿瘤生长情况、转移情况,以期建立更稳定、更接近于人肺癌生长情况的小鼠肺癌原位模型。方法将处于对数生长期的Lewis肺癌细胞混悬于Matrigel中,接种于C57BL/6近交系小鼠左肺内。分别于第4、7、10、13、16天各处死5只小鼠,观察其局部成瘤率、肿瘤生长情况、中位生存期及肿瘤转移情况,并对各阶段小鼠行肺部,肝脏,肾脏,脾脏病理切片检查。结果术后第7天解剖的5只小鼠中,3只小鼠肺上可见小的瘤结节形成,其余2只肺上未见肉眼成瘤,行病理HE染色检查在显微镜下可见2只小鼠肺脏有小的瘤结节形成。术后第10天以后处死的所有小鼠肺上均有肉眼成瘤,术后第13天,所有小鼠肺原位成瘤并伴有血性胸腔积液、胸腔内转移。术后第25天,有1只小鼠出现上述转移的同时还出现了心包膜转移及肾脏远处转移。5只小鼠生存期分别为17 d、20 d、22 d、22 d、25 d,小鼠中位生存期为21.2 d（17~25 d）。成瘤率100%。结论利用Matrigel法成功建立小鼠Lewis肺癌原位模型,稳定性好,成瘤率高,并具有远处转移的特性,更接近于人肺癌的发生、发展过程。     

利用裸鼠建立人泌尿生殖系统肿瘤细胞系

目的建立人泌尿系肿瘤无限细胞系,为泌尿系肿瘤研究提供实验模型.方法无菌取下肿瘤标本后,将标本剪成大小约1.0mm3的组织块,在裸鼠右后肢皮下包埋,当皮下肿瘤块发生明显增殖并长到一定程度后,再行裸鼠体内传代两次,最后取下组织块进行原代培养.培养细胞传代超过20代后按建系标准[2]进行检测.结果共取40例标本,裸鼠体内传代F1代成功6例,F3代成功3例,该3例标本行原代培养后建成3个无限细胞系人肾透明细胞癌RCC-9863,人膀胱癌BC-6,人前列腺癌PC-98106,全部细胞传代1年以上,生长稳定,传代周期固定,其形态结构,分化程度与原发瘤保持一致,染色体形态仍为人类核型.结论裸鼠肿瘤皮下种植法是泌尿系肿瘤建系的一个较好方法.     

A transgenic red fluorescent protein‐expressing nude mouse for color‐coded imaging of the tumor microenvironment

The tumor microenvironment (TME) is critical for tumor growth and progression. We have previously developed color‐coded imaging of the TME using a green fluorescent protein (GFP) transgenic nude mouse as a host. However, most donor sources of cell types appropriate for study in the TME are from mice expressing GFP. Therefore, a nude mouse expressing red fluorescent protein (RFP) would be an appropriate host for transplantation of GFP‐expressing stromal cells as well as double‐labeled cancer cells expressing GFP in the nucleus and RFP in the cytoplasm, thereby creating a three‐color imaging model of the TME. The RFP nude mouse was obtained by crossing non‐transgenic nude mice with the transgenic C57/B6 mouse in which the β‐actin promoter drives RFP (DsRed2) expression in essentially all tissues. In crosses between nu/nu RFP male mice and nu/+ RFP female mice, the embryos fluoresced red. Approximately 50% of the offspring of these mice were RFP nude mice. In the RFP nude mouse, the organs all brightly expressed RFP, including the heart, lungs, spleen, pancreas, esophagus, stomach, duodenum, the male and female reproductive systems; brain and spinal cord; and the circulatory system, including the heart, and major arteries and veins. The skinned skeleton highly expressed RFP. The bone marrow and spleen cells were also RFP positive. GFP‐expressing human cancer cell lines, including HCT‐116‐GFP colon cancer and MDA‐MB‐435‐GFP breast cancer were orthotopically transplanted to the transgenic RFP nude mice. These human tumors grew extensively in the transgenic RFP nude mouse. Dual‐color fluorescence imaging enabled visualization of human tumor–host interaction. The RFP nude mouse model should greatly expand our knowledge of the TME. J. Cell. Biochem. 106: 279–284, 2009. © 2008 Wiley‐Liss, Inc.     

建立小鼠淋巴瘤模型所需环磷酰胺最佳预处理剂量探索

目的:探讨不同剂量环磷酰胺对小鼠肿瘤成模情况的影响,寻找一种简单、有效的建立肿瘤模型的预处理方法。方法:给予BALB/c裸鼠腹腔注射不同剂量环磷酰胺,72小时后再给予小鼠皮下接种淋巴瘤细胞,观察预处理前后小鼠外周血白细胞数量及体重变化情况,以及肿瘤成模率、急性死亡率等。结果:①组1(NS对照组)、组2(100mg/kg×1d)、组3(125mg/kg×1d)、组4(75mg/kg×2d)预处理后体重较处理前无显著性变化,亦无急性死亡情况发生;而组5(125mg/kg×2d)、组6(200mg/kg×2d)、组7(125mg/kg×3d)、组8(250mg/kg×3d)小鼠体重较预处理前明显减轻,且急性死亡率依次为30%、58.3%、50%、75%;②组1和组2小鼠预处理后72小时外周血白细胞数较处理前无明显差异,同时均未成模;而组3、组4、组5、组6、组7、组8小鼠白细胞较预处理前均显著下降,成模率依次为20%、83.3%、60%、33.3%、50%、25%。结论:使用环磷酰胺75mg/kg连续2天腹腔注射的预处理方案,操作简单,成本低廉,通过观察外周血白细胞数和小鼠体重水平等指标即可初步判断建模情况,同时肿瘤成模率高,毒副作用小,是理想的预处理方案。     

5-烯丙基-7-二氟亚甲基白杨素对人肺癌A549细胞裸鼠移植瘤生长的影响

目的研究5-烯丙基-7-二氟亚甲基白杨素（ADFMChR）对人肺癌A549细胞裸鼠移植瘤生长的影响。方法建立人肺癌A549细胞裸鼠移植瘤模型,测定荷人肺癌裸鼠移植瘤的大小和重量,应用免疫组化SP法检测移植瘤组织中PCNA、VEGF、CD31的表达。结果ADFMChR对肺癌移植瘤生长有显著抑制作用（P〈0.01）,5.0、10.0、20.0 mg/（kg.bw）的ADFMChR对移植瘤的瘤重抑制率分别为42.98%,82.31%和89.91%。免疫组化检测结果表明：ADFMChR具有抑制肺腺癌裸鼠移植瘤细胞PCNA、VEGF及CD31蛋白表达作用。结论ADFMChR抑制肺腺癌裸鼠移植瘤的生长作用与其抑制移植瘤细胞PCNA、VEGF以及CD31的蛋白表达相关。     

SW1990和Capan-2红色荧光标记胰腺癌移植肿瘤模型的建立和比较

目的建立稳定表达红色荧光蛋白基因的人胰腺癌细胞系,为体内监测肿瘤的早期生长及抗肿瘤药物的药效评价建立一种新的肿瘤动物模型。方法以Lipofectamine 2000介导chickenβ-actin-RFP-NEO转染人胰腺癌细胞SW1990和Capan-2,经梯度浓度G418筛选获得稳定表达红色荧光蛋白的细胞克隆并扩大培养。BALB/cA-nu裸鼠皮下接种1×106个发光细胞使其成瘤,活体荧光成像系统观察肿瘤的生长情况。结果获得了稳定表达RFP的两种不同的人胰腺癌细胞株,将其接种到裸鼠体内可成瘤,利用活体成像系统观察了肿瘤的生长动态过程,并且SW1990肿瘤细胞的生长速度较Capan-2细胞快。结论用红色荧光蛋白标记的人胰腺癌细胞建立的裸鼠肿瘤模型为胰腺癌的研究和相关药物筛选提供了可进行荧光影像活体、动态分析的动物模型。     

Tumor-targeting amino acid auxotrophic <Emphasis Type="Italic">Salmonella typhimurium</Emphasis>

We have developed an effective bacterial cancer therapy strategy by targeting viable tumor tissue using Salmonella typhimurium auxotrophs that we have generated which grow in viable as well as necrotic areas of tumors. However, the auxotrophy severely restricts growth of these bacteria in normal tissue. The S. typhimurium A1-R mutant, which is auxotrophic for leu-arg and has high anti-tumor virulence, was developed in our laboratory. In vitro, A1-R infects tumor cells and causes nuclear destruction. A1-R was initially used to treat metastatic human prostate and breast tumors that had been orthotopically implanted in nude mice. Forty percent of treated mice were cured completely and survived as long as non-tumor-bearing mice. A1-R administered i.v. to nude mice with primary osteosarcoma and lung metastasis was highly effective, especially against metastasis. A1-R was also targeted to both axillary lymph and popliteal lymph node metastasis of human pancreatic cancer and fibrosarcoma, respectively, as well as lung metastasis of the fibrosarcoma in nude mice. The bacteria were delivered via a lymphatic channel to target the lymph node metastases and systemically via the tail vein to target the lung metastasis. The metastases were cured without the need of chemotherapy or any other treatment. A1-R was administered intratumorally to nude mice with an orthotopically transplanted human pancreatic tumor. The primary pancreatic cancer regressed without additional chemotherapy or any other treatment. A1-R was also effective against pancreatic cancer liver metastasis when administered intrasplenically to nude mice. The approach described here, where bacterial monotherapy effectively treats primary and metastatic tumors, is a significant improvement over previous bacterial tumor-therapy strategies that require combination with toxic chemotherapy. Three promoter clones engineered in S. enterica typhimurium were identified to have enhanced expression in bacteria growing in tumors relative to those growing in the spleen. The expression of therapeutics in Salmonella under the regulation of one or more promoters that are activated preferentially in tumors has the potential to improve the efficacy of Salmonella tumor therapy. Exploitation of the tumor-killing capability of Salmonella has great promise for a new paradigm of cancer therapy.     

Antitumor activity of a novel antisense oligonucleotide against Akt1

The AKT pathway is an important therapeutic target for cancer drug discovery as it functions as a main point for transducing extracellular and intracellular oncogenic signals. Moreover, alternations of the AKT pathway have been found in a wide range of cancers. In the present study, we found that an Akt1 antisense oligonucleotide (Akt1 AO) significantly downregulated the expression of AKT1 at both the mRNA and protein levels and inhibited cellular growth at nanomolar concentrations in various types of human cancer cells. Combined treatment of Akt1 AO with several cytotoxic drugs resulted in an additive growth inhibition of Caki‐1 cells. The in vivo effectiveness of Akt1 AO was determined using two different xenograft nude mouse models. Akt1 AO (30 mg/kg, i.v. every 48 h) significantly inhibited the tumor growth of nude mouse subcutaneously implanted with U251 human glioblastoma cells after 27 days treatment. Akt1 AO (30 mg/kg, i.p continuously via osmotic pump) also significantly inhibited the tumor formation in nude mice implanted with luciferase‐expressing MIA human pancreatic cancer cells (MIA‐Luc) after 14 days of treatment. The luciferase signals from MIA‐Luc cells were reduced or completely abolished after 2 weeks of treatment and the implanted tumors were barely detectable. Our findings suggest that Akt1 AO alone or in combination with other clinically approved anticancer agents should be further explored and progressed into clinical studies as a potential novel therapeutic agent. J. Cell. Biochem. 108: 832–838, 2009. © 2009 Wiley‐Liss, Inc.     

存活素反义寡核苷酸在人甲状腺髓样癌裸鼠模型中的抑瘤作用

目的探讨存活素(survivin)反义寡核苷酸(antisense oligodeoxyribonucleotide,ASODN)在人甲状腺髓样癌裸鼠模型中的抑瘤作用。方法构建12只人甲状腺髓样癌裸鼠模型,随机分配到正常组,SODN(sense oligodeoxyribonucleotide,正义寡核苷酸)组和ASODN组,每组4只。各组每日一次瘤内注射相应处理液,连续7d。治疗结束后1周,杀死全部瘤鼠,切取瘤体,测算瘤重和抑瘤率以评价肿瘤生长情况,瘤组织石腊切片分别行HE、免疫组化(IHC)和DNA末端原位标记法(TUNEL法)染色以评价组织病理构造、survivin与VEGF蛋白表达和细胞凋亡的变化。结果HE染色显示ASODN组具有明显减低的血管新生特点。此外,相对于正常组和SODN组,ASODN组具有显著的survivin和VEGF低表达、高凋亡、低瘤重和高抑瘤率特点(各P<0.01),而正常组与SODN组间未见上述各特点的统计学差异。结论本研究证实针对人甲状腺髓样癌survivin基因沉默治疗能够达到明显抑瘤效果,此效果至少部分通过促进肿瘤调亡和抑制肿瘤血管新生而实现。这提示survivin基因在人甲状腺髓样癌中具有重要意义和其可成为人甲状腺髓样癌基因靶向沉默治疗的重要候选基因之一。     

两种人乳腺癌裸鼠移植模型的建立

目的建立简便的人乳腺癌裸鼠移植模型,并探讨其部分生物学特性。方法采用雌激素受体阴性的MDA-MB-231和SK-BR-3人乳腺癌细胞株,分别接种于10只裸鼠左侧腋窝皮下,移植细胞总数为1×107/只。观察肿块生长情况,第42天处死荷瘤鼠,切除肿块作病理切片。结果 MDA-MB-231接种后第5d在接种部位可见结节,成瘤率为90%（9/10）,接种42 d肿瘤体积426.6±333.8,瘤重0.417±0.276,病理学检查为浸润性导管癌;SK-BR-3接种后第11天在接种部位可见结节,成瘤率为80%（8/10）,接种42 d肿瘤体积357.5±246,瘤重0.325±0.167,病理学检查为浸润性导管癌。结论该方法建立的人乳腺癌裸鼠移植模型,皮下移植方法简单,易于操作,成功率较高,肿瘤可部分保持人乳腺癌生物学特性,为研究人乳腺癌提供了重要工具。     

胡桃醌不同给药途径对人肝癌细胞BEL-7402裸鼠皮下移植瘤生长的影响

目的采用裸鼠皮下移植瘤模型,通过不同给药途径对胡桃醌抗肿瘤活性和毒性进行评价。方法建立人肝癌BEL-7402细胞裸鼠皮下移植瘤模型,通过腹腔注射和局部注射两个给药途径观察胡桃醌抑制肿瘤生长的效果。结果①以600、300和150μg/kg胡桃醌腹腔注射于人肝癌BEL-7402细胞裸鼠皮下移植瘤模型,发现该剂量胡桃醌对肿瘤生长没有明显的影响;NK细胞活性检测发现,600、300μg/kg胡桃醌对裸鼠免疫功能有影响(P均<0.01),150μg/kg胡桃醌则没有影响(P>0.05);与阳性对照组(5-Fu)相比,600μg/kg胡桃醌组NK细胞活性差异无显著性(P>0.05),300和150μg/kg胡桃醌组NK细胞活性差异有显著性(P<0.05,P<0.01),结果提示胡桃醌对小鼠免疫系统有一定的损伤作用。②以4.5、3和1.5 mg/kg胡桃醌腹腔注射于人肝癌BEL-7402细胞裸鼠皮下移植瘤模型,抑瘤率分别为为78.24%、66.57%、48.94%;4.5、3 mg/kg胡桃醌的抑瘤作用可与阳性对照组比拟(P均>0.05)。但4.5 mg/kg胡桃醌组裸鼠出现明显的皮下脂肪减少、消瘦,并有死亡现象。③以pH 7.4和pH 4.0的600、300和150μg/kg胡桃醌人肝癌BEL-7402细胞裸鼠皮下移植瘤模型局部给药,结果发现不同pH(pH 7.4或4.0)600、300μg/kg的胡桃醌局部注射抑瘤作用与阳性对照组(5-Fu)组差异无显著性(P>0.05),而不同pH的150μg/kg胡桃醌抑瘤作用不明显。同一浓度不同pH药物的抑瘤作用差异无显著性(P均>0.05),但pH 4.0的胡桃醌组肿瘤细胞肝转移较少。结论胡桃醌不同给药途径均可抑制人肝癌BEL-7402细胞裸鼠皮下移植瘤的生长,但有一定的毒副作用,药物安全范围较小。     

Curcumin induces apoptosis and inhibits growth of orthotopic human non-small cell lung cancer xenografts

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related mortality. Curcumin is involved in various biological pathways leading to inhibition of NSCLC growth. The purpose of this study was to evaluate the effect of curcumin on expression of nuclear factor κB-related proteins in vitro and in vivo and on growth and metastasis in an intralung tumor mouse model.H1975 NSCLC cells were treated with curcumin (0–50 μM) alone, or combined with gemcitabine or cisplatin. The effects of curcumin were evaluated in cell cultures and in vivo, using ectopic and orthotopic lung tumor mouse models. Twenty mice were randomly selected into two equal groups, one that received AIN-076 control diet and one that received the same food but with the addition of 0.6% curcumin 14 days prior to cell implantation and until the end of the experiment. To generate orthotopic tumor, lung cancer cells in Matrigel were injected percutaneously into the left lung of CD-1 nude mice. Western blot analysis showed that the expressions of IkB, nuclear p65, cyclooxygenase 2 (COX-2) and p-ERK1/2 were down-regulated by curcumin in vitro. Curcumin potentiated the gemcitabine- or cisplatin-mediated antitumor effects. Curcumin reduced COX-2 expression in subcutaneous tumors in vivo and caused a 36% decrease in weight of intralung tumors (P=.048) accompanied by a significant survival rate increase (hazard ratio=2.728, P=.036). Curcumin inhibition of COX-2, p65 expression and ERK1/2 activity in NSCLC cells was associated with decreased survival and increased induction of apoptosis. Curcumin significantly reduced tumor growth of orthotopic human NSCLC xenografts and increased survival of treated athymic mice. To evaluate the role of curcumin in chemoprevention and treatment of NSCLC, further clinical trials are required.     

裸鼠肿瘤动物模型VEGF受体表达及其意义

目的　通过免疫组织化学染色了解flt 1与flk 1 KDR(VEGF的两个高亲和受体 )在人肿瘤细胞皮下接种肿瘤动物模型的血管内皮细胞与肿瘤细胞中的表达。方法　取荷瘤裸鼠皮下接种瘤块 ,漂洗、固定、石蜡连续切片 ,进行两种受体相应免疫组化检测。结果　在 13种荷瘤裸鼠血管内皮细胞及肿瘤细胞中flt 1的阳性率大部分为强阳性或中阳性 ,而只有在荷人胃腺癌MKN 4 5裸鼠的肿瘤细胞中flt 1的阳性率为弱阳性 ,在荷人卵巢癌SKOv3裸鼠的肿瘤细胞中flt 1的表达为阴性。相比较而言 ,在 13种荷瘤裸鼠血管内皮细胞及肿瘤细胞中KDR的阳性率大部分为中阳性或弱阳性 ,并且在荷人肝癌SMMC 772 1裸鼠 ,荷人胃腺癌SPC A1裸鼠 ,荷人高转移肝癌移植瘤裸鼠 ,荷人卵巢癌SKOv3裸鼠的肿瘤细胞中 ,荷人宫颈癌移植瘤裸鼠和荷人胃腺癌MKN 4 5裸鼠的肿瘤细胞中 ,KDR表达为阴性。结论　VEGF受体共同表达于肿瘤血管内皮细胞与肿瘤细胞 ,提示了VEGF与VEGF受体结合作用在肿瘤演化中的重要性 ,为靶向于VEGF受体的基因治疗策略选择裸鼠动物模型提供了参考依据     

人结肠腺癌细胞HCT－8绿色荧光标记肿瘤模型的建立

目的筛选表达绿色荧光蛋白基因的人的单克隆结肠腺癌细胞系,为体内监测肿瘤的早期生长建立一种新的肿瘤动物模型。方法以脂质体2000介导chicken β－acfin-GFP—NEO转染人结肠腺癌细胞HCT－8,经梯度浓度G418筛选获得稳定表达绿色荧光蛋白的细胞克隆并扩大培养。BALB／CA－nu裸鼠皮下接种1×10^6个发光细胞使其成瘤,活体荧光成像系统观察肿瘤的生长情况。结果获得了稳定表达GFP的人结肠腺癌细胞株,将其接种到裸鼠体内可成瘤,利用活体成像系统观察了肿瘤的生长过程,肿瘤的发光随着观察时间的延长而增加。结论绿色荧光蛋白能够在人结肠腺癌细胞HCT-8中长期稳定表达,用绿色荧光蛋白标记的人结肠腺癌细胞HCT-8建立的裸鼠肿瘤模型为进一步研究结肠肿瘤和相应的药物筛选提供了一种简便、可行的新方法。     

Establishment of a Patient-Derived Orthotopic Xenograft (PDOX) Model of HER-2-Positive Cervical Cancer Expressing the Clinical Metastatic Pattern

Squamous cell carcinoma of the cervix, highly prevalent in the developing world, is often metastatic and treatment resistant with no standard treatment protocol. Our laboratory pioneered the patient-derived orthotopic xenograft (PDOX) nude mouse model with the technique of surgical orthotopic implantation (SOI). Unlike subcutaneous transplant patient-derived xenograft (PDX) models, PDOX models metastasize. Most importantly, the metastasis pattern correlates to the patient. In the present report, we describe the development of a PDOX model of HER-2-positive cervical cancer. Metastasis after SOI in nude mice included peritoneal dissemination, liver metastasis, lung metastasis as well as lymph node metastasis reflecting the metastatic pattern in the donor patient. Metastasis was detected in 4 of 6 nude mice with primary tumors. Primary tumors and metastases in the nude mice had histological structures similar to the original tumor and were stained by an anti-HER-2 antibody in the same pattern as the patient’s cancer. The metastatic pattern, histology and HER-2 tumor expression of the patient were thus preserved in the PDOX model. In contrast, subcutaneous transplantation of the patient’s cervical tumors resulted in primary growth but not metastasis.