Bioluminescent human breast cancer cell lines that permit rapid and sensitive in vivo detection of mammary tumors and multiple metastases in immune deficient mice.
Bioluminescent human breast cancer cell lines that permit rapid and sensitive in vivo detection of mammary tumors and multiple metastases in immune deficient mice.
复制标题
DOI：
10.1186/bcr1026
复制
发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Purchio T
中科院分区：
文献类型：
--
作者：
Jenkins DE;Hornig YS;Oei Y;Dusich J;Purchio T
我们的目标是生成基于表达荧光素酶的MDA-MB-231肿瘤细胞的人类乳腺癌异种移植小鼠模型，该模型将提供快速的乳腺肿瘤生长；产生转移到临床相关组织，如淋巴结、肺和骨；并允许敏感的体内检测原发和继发肿瘤部位的生物发光成像。用荧光素酶cDNA转染亲本细胞，分离出稳定表达萤火虫荧光素酶的人MDA-MB-231细胞的两个克隆细胞亚系。每个亚系在体内传代一次或两次，以促进原发肿瘤的生长和增加转移。通过生物发光成像分析得到的表达荧光素酶的D3H1和D3H2LN细胞的长期生物发光稳定性、原发肿瘤生长情况以及远端淋巴结、肺、骨和软组织转移情况。将细胞注射到裸小鼠和裸米色小鼠的乳腺脂肪垫或通过心脏内注射全身给药。转移也通过离体成像和死后组织学分析进行评估。D3H1和D3H2LN细胞系在体内表现出长达4-6个月的肿瘤累积生长时间的长期稳定的荧光素酶表达。生物发光成像量化原发性乳腺脂肪垫肿瘤的发展，并在体内检测早期自发淋巴结转移。与D3H1肿瘤相比，D3H2LN肿瘤自发淋巴结转移的频率增加。通过死后离体成像，我们在D3H2LN乳腺肿瘤小鼠中发现了额外的肺部微转移。随后对淋巴结和肺叶组织切片的组织学评估证实了这些部位的自发肿瘤转移。经心内注射MDA-MB-231-luc肿瘤细胞后，发现肿瘤早期向骨组织、淋巴结、脑及各脏器转移。每周的体内成像数据允许同时对多个部位的转移进行纵向分析。来自样本组织的离体成像数据证实了骨骼和多个软组织肿瘤转移。本研究鉴定了两种新的生物发光MDA-MB-231-luc人乳腺癌细胞系，它们在小鼠中具有增强的肿瘤生长和广泛的转移。它们在目前乳腺癌异种移植模型中的应用为体内抗癌治疗的临床前评估提供了快速和高灵敏度的检测选择。
Our goal was to generate xenograft mouse models of human breast cancer based on luciferase-expressing MDA-MB-231 tumor cells that would provide rapid mammary tumor growth; produce metastasis to clinically relevant tissues such as lymph nodes, lung, and bone; and permit sensitive in vivo detection of both primary and secondary tumor sites by bioluminescent imaging. Two clonal cell sublines of human MDA-MB-231 cells that stably expressed firefly luciferase were isolated following transfection of the parental cells with luciferase cDNA. Each subline was passaged once or twice in vivo to enhance primary tumor growth and to increase metastasis. The resulting luciferase-expressing D3H1 and D3H2LN cells were analyzed for long-term bioluminescent stability, primary tumor growth, and distal metastasis to lymph nodes, lungs, bone and soft tissues by bioluminescent imaging. Cells were injected into the mammary fat pad of nude and nude-beige mice or were delivered systemically via intracardiac injection. Metastasis was also evaluated by ex vivo imaging and histologic analysis postmortem. The D3H1 and D3H2LN cell lines exhibited long-term stable luciferase expression for up to 4–6 months of accumulative tumor growth time in vivo. Bioluminescent imaging quantified primary mammary fat pad tumor development and detected early spontaneous lymph node metastasis in vivo. Increased frequency of spontaneous lymph node metastasis was observed with D3H2LN tumors as compared with D3H1 tumors. With postmortem ex vivo imaging, we detected additional lung micrometastasis in mice with D3H2LN mammary tumors. Subsequent histologic evaluation of tissue sections from lymph nodes and lung lobes confirmed spontaneous tumor metastasis at these sites. Following intracardiac injection of the MDA-MB-231-luc tumor cells, early metastasis to skeletal tissues, lymph nodes, brain and various visceral organs was detected. Weekly in vivo imaging data permitted longitudinal analysis of metastasis at multiple sites simultaneously. Ex vivo imaging data from sampled tissues verified both skeletal and multiple soft tissue tumor metastasis. This study characterized two new bioluminescent MDA-MB-231-luc human breast carcinoma cell lines with enhanced tumor growth and widespread metastasis in mice. Their application to current xenograft models of breast cancer offers rapid and highly sensitive detection options for preclinical assessment of anticancer therapies in vivo.
登录
查看更多内容
DOI：
10.1093/jnci/53.3.661
发表时间：
1974-01-01
期刊：
JOURNAL OF THE NATIONAL CANCER INSTITUTE
影响因子：
--
作者：
CAILLEAU, R;YOUNG, R;REEVES, WJ
通讯作者：
REEVES, WJ
影响因子：
4.8
作者：
Rehemtulla, A;Stegman, LD;Ross, BD
通讯作者：
Ross, BD
影响因子：
6
作者：
Wetterwald, A;van der Pluijm, G;Cecchini, MG
通讯作者：
Cecchini, MG
影响因子：
15.9
作者：
Minn, AJ;Kang, YB;Massagué, J
通讯作者：
Massagué, J
影响因子：
50.3
作者：
Kang, YB;Siegel, PM;Massagué, J
通讯作者：
Massagué, J