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. 2010 Dec;12(12):1041–1053. doi: 10.1593/neo.10916

Figure 8.

Figure 8

Depletion of TFF3 by siRNA or inhibition of TFF3 function by antibody increases anti-estrogen sensitivity in MCF-7 cells and improves tamoxifen sensitivity in tamoxifen-resistant cells. (A and B) Colony formation in soft agar: MCF7-siCK and MCF7-siTFF3 cells were seeded in 0.35% agar and treated with tamoxifen (TAM) and fulvestrant (Fulv) in 10% FBS at the concentrations indicated for 10 days. (C) Three-dimensional Matrigel: MCF7-siCK and MCF7-siTFF3 cells were seeded in 4% Matrigel and treated with vehicle (Veh) or 1 µM tamoxifen or 10 nM fulvestrant for 7 days. (D) Three-dimensional Matrigel: MCF-7 cells were seeded in 4% Matrigel and treated with 1 µM tamoxifen alone or in combination with 500 µg/ml TFF3-pAb or control rabbit IgG for 7 days. (E) TAMS and TAMR cells were cultured in phenol red-free RPMI medium containing 10% CS-FBS for 3 days and then treated with either vehicle or 1 µM tamoxifen for 24 hours. TFF3 expression was determined by semiquantitative RT-PCR and Western blot analysis. (F) TAMS and TAMR cells were seeded in 4% Matrigel in 10% CS-FBS medium and treated with 1 µM tamoxifen alone or in combination with 500 µg/ml TFF3-pAb or control rabbit IgG for 7 days. *P < .05, **P < .01, ***P < .001 compared with vector control group for the same treatment (A, B, and C) or compared with control IgG and vehicle-treated group (D and F).