association of invasion-promoting tenascin-c additional domains with breast cancers in young women协会invasion-promoting tenascin-c额外的域在年轻女性乳腺癌.pdfVIP

Guttery et al. Breast Cancer Research 2010, 12:R57 /content/12/4/R57 RESEARCH ARTICLE Open Access Association of invasion-promoting tenascin-C additional domains with breast cancers in young women 1 1 2 2 1 1 David S Guttery , Rachael A Hancox , Kellie T Mulligan , Simon Hughes , Sinead M Lambe , J Howard Pringle , Rosemary A Walker1 2 1* , J Louise Jones , Jacqueline A Shaw Abstract Introduction: Tenascin-C (TNC) is a large extracellular matrix glycoprotein that shows prominent stromal expression in many solid tumours. The profile of isoforms expressed differs between cancers and normal breast, with the two additional domains AD1 and AD2 considered to be tumour associated. The aim of the present study was to investigate expression of AD1 and AD2 in normal, benign and malignant breast tissue to determine their relationship with tumour characteristics and to perform in vitro functional assays to investigate the role of AD1 in tumour cell invasion and growth. Methods: Expression of AD1 and AD2 was related to hypoxanthine phosphoribosyltransferase 1 as a housekeeping gene in breast tissue using quantitative RT-PCR, and the results were related to clinicopathological features of the tumours. Constructs overexpressing an AD1-containing isoform (TNC-14/AD1/16) were transiently transfected into breast carcinoma cell lines (MCF-7, T-47 D, ZR-75-1, MDA-MB-231 and GI-101) to assess the effect in vitro on invasion and growth. Statistical analysis was performed using a nonparametric Mann-Whitney test for comparison of clinicopathological features with levels of TNC expression and using Jonckheere-Terpstra trend analysis for associa