An oestrogen-dependent model of breast cancer created by transformation of normal human mammary epithelial cells.
An oestrogen-dependent model of breast cancer created by transformation of normal human mammary epithelial cells.
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DOI：
10.1186/bcr1734
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
Iggo RD
中科院分区：
文献类型：
--
作者：
Duss S;André S;Nicoulaz AL;Fiche M;Bonnefoi H;Brisken C;Iggo RD
大约70%的乳腺癌表达雌激素受体α (ESR1/ERα)，并且在生长过程中依赖于雌激素。与er α阳性肿瘤细胞的高增殖性相反，正常乳腺组织中er α阳性细胞很少增殖。由于在体外培养正常人乳腺上皮细胞（HMECs）时，ERα的表达会迅速丧失，因此由HMECs衍生的乳腺癌模型是ERα阴性的。目前只有肿瘤细胞系可用来模拟er α阳性疾病。为了建立ERα阳性的乳腺癌模型，我们强迫来自缩乳成形术组织的正常hmec表达ERα和其他相关乳腺癌基因。根据乳腺癌微阵列数据选择候选基因，并将其克隆到慢病毒载体中。用慢病毒颗粒感染乳房缩小成形术组织制备的原代hmec。采用Western blotting、免疫荧光显微镜、微阵列分析、生长曲线、核型和SNP芯片分析对感染的hmec进行表征。原位注射NOD/SCID小鼠腹股沟乳腺，检测修饰的HMECs的致瘤性。细胞被荧光蛋白标记，以便在脂肪垫中可视化。通过乳腺荧光显微镜和染色组织切片病理分析，观察移植物的生长情况。通过雌激素拮抗剂氟维司汀治疗来评估肿瘤生长的雌激素依赖性。er α阳性肿瘤的微阵列分析显示，它们通常过表达Polycomb-group基因BMI1。慢病毒通过ERα、BMI1、TERT和MYC进行转导，使原代hmec能够以雌激素依赖的方式在体外扩增。将这些细胞原位移植到NOD/SCID小鼠的乳腺中，可形成er α阳性肿瘤，并转移到多个器官。这些细胞保持TP53的野生型、二倍体和遗传稳定。体内肿瘤的生长和体外肿瘤细胞的增殖都依赖于雌激素。我们已经建立了一个基于正常hmec的er α阳性人类乳腺癌的遗传定义模型，该模型有可能在小鼠中模拟人类雌激素依赖性乳腺癌，并使研究肿瘤发生和转移的机制成为可能。
About 70% of breast cancers express oestrogen receptor α (ESR1/ERα) and are oestrogen-dependent for growth. In contrast with the highly proliferative nature of ERα-positive tumour cells, ERα-positive cells in normal breast tissue rarely proliferate. Because ERα expression is rapidly lost when normal human mammary epithelial cells (HMECs) are grown in vitro, breast cancer models derived from HMECs are ERα-negative. Currently only tumour cell lines are available to model ERα-positive disease. To create an ERα-positive breast cancer model, we have forced normal HMECs derived from reduction mammoplasty tissue to express ERα in combination with other relevant breast cancer genes. Candidate genes were selected based on breast cancer microarray data and cloned into lentiviral vectors. Primary HMECs prepared from reduction mammoplasty tissue were infected with lentiviral particles. Infected HMECs were characterised by Western blotting, immunofluorescence microscopy, microarray analysis, growth curves, karyotyping and SNP chip analysis. The tumorigenicity of the modified HMECs was tested after orthotopic injection into the inguinal mammary glands of NOD/SCID mice. Cells were marked with a fluorescent protein to allow visualisation in the fat pad. The growth of the graft was analysed by fluorescence microscopy of the mammary glands and pathological analysis of stained tissue sections. Oestrogen dependence of tumour growth was assessed by treatment with the oestrogen antagonist fulvestrant. Microarray analysis of ERα-positive tumours reveals that they commonly overexpress the Polycomb-group gene BMI1. Lentiviral transduction with ERα, BMI1, TERT and MYC allows primary HMECs to be expanded in vitro in an oestrogen-dependent manner. Orthotopic xenografting of these cells into the mammary glands of NOD/SCID mice results in the formation of ERα-positive tumours that metastasise to multiple organs. The cells remain wild type for TP53, diploid and genetically stable. In vivo tumour growth and in vitro proliferation of cells explanted from tumours are dependent on oestrogen. We have created a genetically defined model of ERα-positive human breast cancer based on normal HMECs that has the potential to model human oestrogen-dependent breast cancer in a mouse and enables the study of mechanisms involved in tumorigenesis and metastasis.
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影响因子：
10.5
作者：
Dontu, G;Abdallah, WM;Wicha, MS
通讯作者：
Wicha, MS
DOI：
10.1073/pnas.0401064101
发表时间：
2004-04-06
影响因子：
11.1
作者：
Kuperwasser, C;Chavarria, T;Weinberg, RA
通讯作者：
Weinberg, RA
影响因子：
8
作者：
Farmer, P;Bonnefoi, H;Iggo, R
通讯作者：
Iggo, R
影响因子：
5.4
作者：
Dull, T;Zufferey, R;Naldini, L
通讯作者：
Naldini, L
影响因子：
64.5
作者：
Métivier, R;Penot, G;Gannon, F
通讯作者：
Gannon, F