Investigation of three new mouse mammary tumor cell lines as models for transforming growth factor (TGF)-β and Neu pathway signaling studies:: identification of a novel model for TGF-β-induced epithelial-to-mesenchymal transition

被引:16
作者
Lenferink, AE [1 ]
Magoon, J [1 ]
Cantin, C [1 ]
O'Connor-McCourt, MD [1 ]
机构
[1] Natl Res Council Canada, Biotechnol Res Inst, Receptor Signaling & Proteom Grp, Montreal, PQ H4P 2R2, Canada
关键词
murine mammary epithelial cell lines; TGF-beta; epithelial-to-mesenchymal transition;
D O I
10.1186/bcr907
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Introduction This report describes the isolation and characterization of three new murine mammary epithelial cell lines derived from mammary tumors from MMTV (mouse mammary tumor virus)/activated Neu + TbetaRII-AS (transforming growth factor [TGF]-beta type II receptor antisense RNA) bigenic mice (BRI-JM01 and BRI-JM05 cell lines) and MMTV/activated Neu transgenic mice (BRI-JM04 cell line). Methods The BRI-JM01, BRI-JM04, and BRI-JM05 cell lines were analyzed for transgene expression, their general growth characteristics, and their sensitivities to several growth factors from the epidermal growth factor (EGF) and TGF-beta families (recombinant human EGF, heregulin-beta(1) and TGF-beta(1)). The BRI-JM01 cells were observed to undergo a striking morphologic change in response to TGF-beta(1), and they were therefore further investigated for their ability to undergo a TGF-beta-induced epithelial-to-mesenchymal transition (EMT) using motility assays and immunofluorescence microscopy. Results We found that two of the three cell lines (BRI-JM04 and BRI-JM05) express the Neu transgene, whereas, unexpectedly, both of the cell lines that were established from MMTV/activated Neu + TbetaRII-AS bigenic tumors (BRI-JM01 and BRI-JM05) do not express the TbetaRII-AS transgene. The cuboidal BRI-JM01 cells exhibit a short doubling time and are able to form confluent monolayers. The BRI-JM04 and BRI-JM05 cell lines are morphologically much less uniform, grow at a much slower rate, and do not form confluent monolayers. Only the BRI-JM05 cells can form colonies in soft agar. In contrast, all three cell lines form colonies in Matrigel, although the BRI-JM04 and BRI-JM05 cell lines do so more efficiently than the BRI-JM01 cell line. All three cell lines express the cell surface marker E-cadherin, confirming their epithelial character. Proliferation assays showed that the three cell lines respond differently to recombinant human EGF and heregulin-beta(1), and that all are growth inhibited by TGF-beta(1), but that only the BRI-JM01 cell line undergoes an EMT and exhibits increased motility upon TGF-beta(1) treatment. Conclusion We suggest that the BRI-JM04 and BRI-JM05 cell lines can be used to investigate Neu oncogene driven mammary tumorigenesis, whereas the BRI-JM01 cell line will be useful for studying TGF-beta(1)-induced EMT.
引用
收藏
页码:R514 / R530
页数:17
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