Cancer associated fibroblasts stimulated by transforming growth factor beta1 (TGF-beta 1) increase invasion rate of tumor cells: a population study.
Cancer associated fibroblasts (CAFs) are believed to promote tumor growth and progression. Our objective was to measure the effect of TGF-beta1 on fibroblasts isolated from invasive breast cancer patients. Fibroblasts were isolated from tissue obtained at surgery from patients with invasive breast cancer (CAF; n = 28) or normal reduction mammoplasty patients (normal; n = 10). Myofibroblast activation was measured by counting cells immunostained for smooth muscle alpha actin (ACTA2) in cultures +/- TGF-beta 1. Conditioned media (CM) was collected for invasion assays and RNA was isolated from cultures incubated in media +/- TGF-beta1 for 24 h. Q-PCR was used to measure expression of cyclin D1, fibronectin, laminin, collagen I, urokinase, stromelysin-1, and ACTA2 genes. Invasion rate was measured in chambers plated with MDA-MB-231 cells and exposed to CM in the bottom chamber; the number of cells that invaded into the bottom chamber was counted. Wilcox Rank Sum tests were used to evaluate differences in CAFs and normal fibroblasts and the effect of TGF-beta 1. There was no difference in percent myofibroblasts or invasion rate between normal and CAF cultures. However, TGF-beta1 significantly increased the percent of myofibroblasts (P < 0.01) and invasion rate (P = 0.02) in CAF cultures. Stromelysin-1 expression was significantly higher in normal versus CAF cultures (P < 0.01). TGF-beta 1 significantly increased ACTA2 expression in both normal and CAF cultures (P < 0.01). Expression of fibronectin and laminin was significantly increased by TGF-beta in CAF cultures (P < 0.01). CAFs were measurably different from normal fibroblasts in response to TGF-beta 1, suggesting that TGF-beta stimulates changes in CAFs that foster tumor invasion.
Department of Animal Science, Michigan State University, B290 Anthony Hall, East Lansing, MI 48824, USA. ande1218@msu.edu
Theresa M
TM
Casey
Jonathan
J
Eneman
Abigail
A
Crocker
Jeffrey
J
White
Joseph
J
Tessitore
Mary
M
Stanley
Seth
S
Harlow
Janice Y
JY
Bunn
Donald
D
Weaver
Hyman
H
Muss
Karen
K
Plaut
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000473:pathology*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D019278:Matrix Metalloproteinase 3 / Q000032:analysis; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics
D053773:Transforming Growth Factor beta1; D014568:Urokinase-Type Plasminogen Activator; D019278:Matrix Metalloproteinase 3