Premium
The Smad3 transgene product restores radiosensitivity and migratory capacity to Smad3−/− clonal bone marrow stromal cell lines
Author(s) -
Greenberger Joel S,
Cao Shaonan,
Zhang Xichen,
Greenberger Emily E,
Goff Julie P,
Wang Hong,
Bahnson Al,
Shields Donna,
Franicola Darcy,
Epperly Michael W
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1328-b
Title The Smad3 Transgene Product Restores Radiosensitivity and Migratory Capacity to Smad3−/ − Clonal Bone Marrow Stromal Cell Lines An intact Smad3 gene product is critical for TGFB function. We established Smad3−/ − and Smad3+/+ long term bone marrow cultures (LTBMCs) and isolated clonal bone marrow stromal cell lines from each. Smad3−/ − cells had a faster cell doubling time (24 hours compared to 48 hours) and increased saturation density compared to +/+ cells (15.3 ± 1.0 x 10 5 cells/25 mm 2 flask compared to 3.8 ± 0.1 x 10 5 , p = 0.003). Plating efficiency was similar (18.3 ± 2.7 compared to 15.5 ± 1.7, p = 0.417). Cells of a subclone expressing the transgene mRNA, designated Smad3−/ − (3), were established. Tissue culture wells of 100 cells per well were followed for 5 days tracking. Smad3+/+ cells migrated significantly faster. (The average velocities were 0.62 μm/min for Smad3+/+ and 0.36 μm/min for Smad3−/ −, p<0.0001). Average daily velocities for Smad3+/+ 0.51, 0.51, 0.52, 0.72, 0.91 μm/min, and for Smad3−/ − 0.28, 0.38, 0.41, 0.37, 0.35 μm/min. The 5 p‐values comparing these cell lines were all <0.0001. Smad3+/+ and Smad3−/ − (3) cells were significantly more radiosensitive than Smad3−/ − cells. Thus, concordance of radioresistance and decreased migratory capacity follows loss of Smad3 gene product.