Quantification of epithelial cell differentiation in mammary glands and carcinomas from DMBA- and MNU-exposed rats.
Quantification of epithelial cell differentiation in mammary glands and carcinomas from DMBA- and MNU-exposed rats.
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DOI：
10.1371/journal.pone.0026145
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Gould MN
中科院分区：
文献类型：
--
作者：
Sharma D;Smits BM;Eichelberg MR;Meilahn AL;Muelbl MJ;Haag JD;Gould MN
大鼠乳腺癌模型已广泛用于研究乳腺癌的发生、进展、预防和干预。然而，未经治疗和暴露于致癌物的大鼠乳腺上皮细胞分化的定量分子数据是有限的。在这里，我们使用针对细胞表面蛋白 CD24、CD29、CD31、CD45、CD49f、CD61、花生凝集素和 Thy-1、细胞内蛋白 CK14、CK19 和 FAK 的抗体以及鬼笔环肽和 Hoechst 染色，通过多色流式细胞术描述大鼠乳腺上皮细胞 (RMEC) 的表征。我们鉴定了管腔和基底/肌上皮群体以及活跃分裂的 RMEC。在易患乳腺癌的近交大鼠中，我们量化了暴露于乳腺癌致癌物 DMBA 和 MNU 后 1、2 和 4 周时 RMEC 群体分化的变化。 DMBA 暴露不会改变基底细胞或管腔细胞的百分比，但上调 CD49f（整合素 α6）表达并增加细胞周期活性。 MNU 暴露导致管腔/基础比率暂时中断，并且 CD49f 没有上调。当比较 DMBA 或 MNU 诱导的乳腺癌时，RMEC 分化谱无法区分。与未经治疗的大鼠乳腺相比，这些癌症显示 CD29（整合素 β1）和 CD49f 表达上调，FAK（粘着斑激酶）激活增加（尤其是在 CD29hi 群体中），并且 CD61（整合素 β3）表达减少。这项研究提供了对 RMEC 分化背后的蛋白质表达表型的定量见解。结果强调了 DMBA 和 MNU 暴露的不同 RMEC 分化病因，而产生的癌症具有相似的 RMEC 分化特征。该方法和数据将增强大鼠乳腺癌模型在乳腺癌中上皮细胞分化作用的研究中的作用。
Rat mammary carcinogenesis models have been used extensively to study breast cancer initiation, progression, prevention, and intervention. Nevertheless, quantitative molecular data on epithelial cell differentiation in mammary glands of untreated and carcinogen-exposed rats is limited. Here, we describe the characterization of rat mammary epithelial cells (RMECs) by multicolor flow cytometry using antibodies against cell surface proteins CD24, CD29, CD31, CD45, CD49f, CD61, Peanut Lectin, and Thy-1, intracellular proteins CK14, CK19, and FAK, along with phalloidin and Hoechst staining. We identified the luminal and basal/myoepithelial populations and actively dividing RMECs. In inbred rats susceptible to mammary carcinoma development, we quantified the changes in differentiation of the RMEC populations at 1, 2, and 4 weeks after exposure to mammary carcinogens DMBA and MNU. DMBA exposure did not alter the percentage of basal or luminal cells, but upregulated CD49f (Integrin α6) expression and increased cell cycle activity. MNU exposure resulted in a temporary disruption of the luminal/basal ratio and no CD49f upregulation. When comparing DMBA- or MNU-induced mammary carcinomas, the RMEC differentiation profiles are indistinguishable. The carcinomas compared with mammary glands from untreated rats, showed upregulation of CD29 (Integrin β1) and CD49f expression, increased FAK (focal adhesion kinase) activation especially in the CD29hi population, and decreased CD61 (Integrin β3) expression. This study provides quantitative insight into the protein expression phenotypes underlying RMEC differentiation. The results highlight distinct RMEC differentiation etiologies of DMBA and MNU exposure, while the resulting carcinomas have similar RMEC differentiation profiles. The methodology and data will enhance rat mammary carcinogenesis models in the study of the role of epithelial cell differentiation in breast cancer.
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影响因子：
4.6
作者：
Guan, Jun-Lin
通讯作者：
Guan, Jun-Lin
影响因子：
21.3
作者：
Taddei, Ilaria;Deugnier, Marie-Ange;Glukhova, Marina A.
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Glukhova, Marina A.
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作者：
RUDLAND, PS
通讯作者：
RUDLAND, PS
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作者：
KIM, ND;CLIFTON, KH
通讯作者：
CLIFTON, KH
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作者：
Papaconstantinou, AD;Shanmugam, I;Snyderwine, EG
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