Characterization of human breast cancer epithelial cells (HBCEC) derived from long term cultured biopsies.
Characterization of human breast cancer epithelial cells (HBCEC) derived from long term cultured biopsies.
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DOI：
10.1186/1756-9966-28-127
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发表时间：
2009-09-14
期刊：
影响因子：
--
通讯作者：
Bertram C
中科院分区：
文献类型：
--
作者：
Hass R;Bertram C
为了更个性化的治疗方法，我们探索了一种无蛋白酶的方法来培养来自乳腺癌活检组织的原代细胞。来自手术后的乳腺癌患者的肿瘤组织在没有酶消化的情况下离体培养超过一年，并显示粘附和增殖的原代细胞群的连续生长。这些人乳腺癌衍生的上皮细胞（HBCEC）的免疫荧光染色和流式细胞术定量显示几乎完全细胞角蛋白表达细胞。原代HBCEC长期肿瘤培养（超过476天）期间的表面标志物分析表明CD 24、CD 44和MUC 1（CD 227）的显着表达。根据衰老标志物，在培养722天后，在原发性肿瘤来源的HBCEC群体中，衰老相关β-半乳糖苷酶的表达显示很少（如果有的话）阳性染色，而大多数正常人乳腺上皮细胞（HMEC）在培养32天后已经显示出衰老细胞。在这种情况下，HBCEC群体来自肿瘤培养后152天和308天，分别表现出显着的端粒酶活性，表明连续的增殖能力。几种化疗化合物及其组合的治疗显示了不同乳腺癌患者HBCEC之间的不同细胞毒性作用，表明这些肿瘤来源的原代细胞的个性化反应。原发性HBCEC的无蛋白酶生长提供了一种患者特异性方法来优化个性化设计的癌症治疗。此外，来自长期乳腺肿瘤组织培养物的HBCEC通过完整的ECM形成和稳定的细胞表面蛋白表达而类似于肿瘤细胞样性质，提供了可重现的筛选平台以鉴定新的生物标志物并在个体肿瘤样品中测试新的治疗剂。
For a more individualized therapeutic approach we explored a protease-free method to culture primary cells from breast cancer biopsies. Tumor tissue from breast cancer patients after surgery was cultured ex vivo without enzymatic digestion for more than one year and revealed the continuous outgrowth of adherent and proliferating primary cell populations. Immunofluorescence staining of these human breast cancer-derived epithelial cells (HBCEC) and quantification by flow cytometry revealed nearly exclusively cytokeratin-expressing cells. Analysis of surface markers during long term tumor culture of primary HBCEC (more than 476d) demonstrated a prominent expression of CD24, CD44 and MUC1 (CD227). According to aging markers, expression of senescence-associated β-galactosidase revealed little if any positive staining in a primary tumor-derived HBCEC population after 722d in culture, whereas the majority of normal human mammary epithelial cells (HMEC) demonstrated senescent cells already after a culture period of 32d. In this context, HBCEC populations derived from a tumor culture after 152d and 308d, respectively, exhibited a significant telomerase activity, suggesting continuous proliferative capacity. Treatment with several chemotherapeutic compounds and their combinations revealed distinct cytotoxic effects among HBCEC from different breast cancer patients, indicating an individualized response of these tumor-derived primary cells. The protease-free outgrowth of primary HBCEC offers a patient-specific approach to optimize an individually-designed cancer therapy. Moreover, HBCEC from long term breast tumor tissue cultures resemble tumor cell-like properties by an intact ECM formation and a stable cell surface protein expression providing a reproducible screening platform to identify new biomarkers and to test new therapeutics in individual tumor samples.
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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：
10.1186/bcr1673
发表时间：
2007
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Fillmore C;Kuperwasser C
通讯作者：
Kuperwasser C
影响因子：
7.5
作者：
Lim, SC
通讯作者：
Lim, SC
影响因子：
3.9
作者：
Bertram, Catharina;Hass, Ralf
通讯作者：
Hass, Ralf
影响因子：
2.9
作者：
FRANKE, WW;SCHMID, E;WEBER, K
通讯作者：
WEBER, K