Generation of tumor-initiating cells by exogenous delivery of OCT4 transcription factor.
Generation of tumor-initiating cells by exogenous delivery of OCT4 transcription factor.
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DOI：
10.1186/bcr3019
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发表时间：
2011-09-27
期刊：
影响因子：
--
通讯作者：
Blancafort P
中科院分区：
文献类型：
--
作者：
Beltran AS;Rivenbark AG;Richardson BT;Yuan X;Quian H;Hunt JP;Zimmerman E;Graves LM;Blancafort P
肿瘤起始细胞（TIC）因其在肿瘤病因学、维持和对治疗的抗性中的作用而被广泛研究。TIC的分离受到了限制，因为在起源组织中缺乏这种群体，并且因为表征这些细胞的分子特征还没有很好地理解。在此，我们描述了通过在原代乳腺细胞制剂中异位表达OCT 4转录因子（TF）来产生TIC样细胞系。OCT 4 cDNA在来自乳房缩小术供体的四种不同的原代人乳腺上皮（HMEC）乳腺细胞制剂中过表达。OCT 4转导的乳腺细胞（OTBC）在自我更新条件下（人胚胎干细胞培养基中的饲养层培养物）产生集落（频率约为0.01%）。进行分化测定、免疫荧光、免疫组织化学和流式细胞术以研究OTBC的细胞来源。将OTBC的系列稀释液注射到裸鼠中以解决其致瘤能力。在OTBC中进行基因表达微阵列，并通过敲低实验研究OCT 4下游靶点在维持自我更新中的作用。OTBC克服衰老，过表达端粒酶，下调p16 INK 4A。在分化条件下，OTBC以低频率产生肌上皮细胞和管腔细胞两者的群体，表明一些OTBC的起源细胞是双能干细胞。在裸鼠中注射OTBC产生具有定植能力的低分化乳腺癌。OTBC系的基因表达微阵列揭示了在乳腺癌的密蛋白-低分子亚型中过度表达的基因特征。最后，siRNA介导的OCT 4或OCT 4下游胚胎靶点（如NANOG和ZIC 1）的敲低抑制了OTBC自我更新的能力。在正常乳腺制备物中转导OCT 4导致产生具有肿瘤起始和定殖能力的细胞系。这些细胞在裸鼠中发展为高级别、低分化的乳腺癌。OTBC的全基因组分析概述了一种胚胎TF回路，该回路可在TIC中起作用，导致癌基因上调和肿瘤抑制功能丧失。这些OTBC代表了用于在低密蛋白肿瘤中发现新型致癌靶点的患者特异性模型系统。
Tumor-initiating cells (TIC) are being extensively studied for their role in tumor etiology, maintenance and resistance to treatment. The isolation of TICs has been limited by the scarcity of this population in the tissue of origin and because the molecular signatures that characterize these cells are not well understood. Herein, we describe the generation of TIC-like cell lines by ectopic expression of the OCT4 transcription factor (TF) in primary breast cell preparations. OCT4 cDNA was over-expressed in four different primary human mammary epithelial (HMEC) breast cell preparations from reduction mammoplasty donors. OCT4-transduced breast cells (OTBCs) generated colonies (frequency ~0.01%) in self-renewal conditions (feeder cultures in human embryonic stem cell media). Differentiation assays, immunofluorescence, immunohistochemistry, and flow cytometry were performed to investigate the cell of origin of OTBCs. Serial dilutions of OTBCs were injected in nude mice to address their tumorigenic capabilities. Gene expression microarrays were performed in OTBCs, and the role of downstream targets of OCT4 in maintaining self-renewal was investigated by knock-down experiments. OTBCs overcame senescence, overexpressed telomerase, and down-regulated p16INK4A. In differentiation conditions, OTBCs generated populations of both myoepithelial and luminal cells at low frequency, suggesting that the cell of origin of some OTBCs was a bi-potent stem cell. Injection of OTBCs in nude mice generated poorly differentiated breast carcinomas with colonization capabilities. Gene expression microarrays of OTBC lines revealed a gene signature that was over-represented in the claudin-low molecular subtype of breast cancer. Lastly, siRNA-mediated knockdown of OCT4 or downstream embryonic targets of OCT4, such as NANOG and ZIC1, suppressed the ability of OTBCs to self-renew. Transduction of OCT4 in normal breast preparations led to the generation of cell lines possessing tumor-initiating and colonization capabilities. These cells developed high-grade, poorly differentiated breast carcinomas in nude mice. Genome-wide analysis of OTBCs outlined an embryonic TF circuitry that could be operative in TICs, resulting in up-regulation of oncogenes and loss of tumor suppressive functions. These OTBCs represent a patient-specific model system for the discovery of novel oncogenic targets in claudin-low tumors.
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DOI：
10.1158/1055-9965.epi-08-0821
发表时间：
2009-10
期刊：
Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology
影响因子：
--
作者：
Hoque MO;Prencipe M;Poeta ML;Barbano R;Valori VM;Copetti M;Gallo AP;Brait M;Maiello E;Apicella A;Rossiello R;Zito F;Stefania T;Paradiso A;Carella M;Dallapiccola B;Murgo R;Carosi I;Bisceglia M;Fazio VM;Sidransky D;Parrella P
通讯作者：
Parrella P
影响因子：
10.5
作者：
Dontu, G;Abdallah, WM;Wicha, MS
通讯作者：
Wicha, MS
影响因子：
7.4
作者：
Fillmore, Christine M.;Kuperwasser, Charlotte
通讯作者：
Kuperwasser, Charlotte
影响因子：
11.2
作者：
Charafe-Jauffret E;Ginestier C;Iovino F;Wicinski J;Cervera N;Finetti P;Hur MH;Diebel ME;Monville F;Dutcher J;Brown M;Viens P;Xerri L;Bertucci F;Stassi G;Dontu G;Birnbaum D;Wicha MS
通讯作者：
Wicha MS
影响因子：
12.3
作者：
Herschkowitz JI;Simin K;Weigman VJ;Mikaelian I;Usary J;Hu Z;Rasmussen KE;Jones LP;Assefnia S;Chandrasekharan S;Backlund MG;Yin Y;Khramtsov AI;Bastein R;Quackenbush J;Glazer RI;Brown PH;Green JE;Kopelovich L;Furth PA;Palazzo JP;Olopade OI;Bernard PS;Churchill GA;Van Dyke T;Perou CM
通讯作者：
Perou CM