New reagents for improved in vitro and in vivo examination of TGF-beta signalling

Rodney B Luwor, Bo Wang, Thao Nheu, Josephine Iaria, Evelyn Tsantikos, Margaret Hibbs, Oliver M Sieber, Hong-Jian Zhu

Research output: Contribution to journalArticleOther

8 Citations (Scopus)

Abstract

Transforming growth factor-beta (TGF-beta) signalling controls many aspects of cell behaviour and is implicated as a key regulator in tumour formation and progression. However, evaluating levels of active TGF-beta in culture medium or patient plasma and gaining definitive information regarding the activity of downstream substrates such as Sma- and Mad-related protein 3 (Smad3) in vivo with accuracy and sensitivity has been problematic. Therefore, to overcome these technical issues we have created a NIH3T3 cell line with stable pCAGA12-luc expression that can now be utilised to detect TGF-beta activity with high sensitivity. In addition, we have created an adenoviral Smad3 luciferase reporter construct pAd.CAGA12-luc to successfully infect cells for in vitro assays, or prior to injection into mice and used to measure transcriptional activity in vivo. Thus, the NIH3T3-pCAGA12-luc cell line and the pAd.CAGA12-luc adenovirus will be extremely useful tools to measure TGF-beta signalling activity with far greater efficiency and reliability compared to original and currently used reagents.
Original languageEnglish
Pages (from-to)211 - 218
Number of pages8
JournalGrowth Factors
Volume29
Issue number5
DOIs
Publication statusPublished - 2011

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