当前位置： SCI文献检索 > BREAST CANCER RESEARCH期刊下所有文献 > A novel approach for the generation of genetically modified mammary epithelial cell cultures yields new insights into TGFβ signaling in the mammary gland.

A novel approach for the generation of genetically modified mammary epithelial cell cultures yields new insights into TGFβ signaling in the mammary gland.

Abstract：

INTRODUCTION:Molecular dissection of the signaling pathways that underlie complex biological responses in the mammary epithelium is limited by the difficulty of propagating large numbers of mouse mammary epithelial cells, and by the inability of ribonucleic acid interference (RNAi)-based knockdown approaches to fully ablate gene function. Here we describe a method for the generation of conditionally immortalized mammary epithelial cells with defined genetic defects, and we show how such cells can be used to investigate complex signal transduction processes using the transforming growth factor beta (TGFβ/Smad pathway as an example. METHODS:We intercrossed the previously described H-2Kb-tsA58 transgenic mouse (Immortomouse) which expresses a temperature-sensitive mutant of the simian virus-40 large T-antigen (tsTAg), with mice of differing Smad genotypes. A panel of conditionally immortalized mammary epithelial cell (IMEC) cultures were derived from the virgin mammary glands of offspring of these crosses and used to assess the Smad dependency of different biological responses to TGFβ. RESULTS:IMECs could be propagated indefinitely at permissive temperatures and had a stable epithelial phenotype, resembling primary mammary epithelial cells with respect to several criteria, including responsiveness to TGFβ. Using this panel of cells, we demonstrated that Smad3, but not Smad2, is necessary for TGFβ-induced apoptotic, growth inhibitory and EMT responses, whereas either Smad can support TGFβ-induced invasion as long as a threshold level of total Smad is exceeded. CONCLUSIONS:This work demonstrates the practicality and utility of generating conditionally immortalized mammary epithelial cell lines from genetically modified Immortomice for detailed investigation of complex signaling pathways in the mammary epithelium.

journal_name

Breast Cancer Res

journal_title

Breast cancer research : BCR

authors

Kohn EA,Du Z,Sato M,Van Schyndle CM,Welsh MA,Yang YA,Stuelten CH,Tang B,Ju W,Bottinger EP,Wakefield LM

doi

10.1186/bcr2728

subject

Has Abstract

pub_date

2010-01-01 00:00:00

pages

R83

issue

eissn

1465-5411

issn

1465-542X

pii

bcr2728

journal_volume

pub_type

杂志文章

在线工具

A novel approach for the generation of genetically modified mammary epithelial cell cultures yields new insights into TGFβ signaling in the mammary gland.