title: New reagents for improved in vitro and in vivo examination of TGF-β signalling.
abstract: Transforming growth factor-β (TGF-β) 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-β 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 pCAGA(12)-luc expression that can now be utilised to detect TGF-β activity with high sensitivity. In addition, we have created an adenoviral Smad3 luciferase reporter construct pAd.CAGA(12)-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-pCAGA(12)-luc cell line and the pAd.CAGA(12)-luc adenovirus will be extremely useful tools to measure TGF-β signalling activity with far greater efficiency and reliability compared to original and currently used reagents.
authors.affiliation: Department of Surgery, University of Melbourne, Parkville, Victoria 3050, Australia.
authors.forename: Rodney B
authors.initials: RB
authors.lastname: Luwor
authors.forename: Bo
authors.initials: B
authors.lastname: Wang
authors.forename: Thao V
authors.initials: TV
authors.lastname: Nheu
authors.forename: Josephine
authors.initials: J
authors.lastname: Iaria
authors.forename: Evelyn
authors.initials: E
authors.lastname: Tsantikos
authors.forename: Margaret L
authors.initials: ML
authors.lastname: Hibbs
authors.forename: Oliver M
authors.initials: OM
authors.lastname: Sieber
authors.forename: Hong-Jian
authors.initials: HJ
authors.lastname: Zhu
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D045744:Cell Line, Tumor; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D051900:Smad3 Protein / Q000235:genetics; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: D051900:Smad3 Protein; D016212:Transforming Growth Factor beta