Objective To study the changes of mRNA expression levels of long non-coding RNA activated by transforming growth factor β (lncRNA-ATB), collagen type Ⅰ (Col I), collagen type Ⅲ (Col Ⅲ) and fibronectin (FN) in lung fibroblasts MRC-5 stimulated by transforming growth factor (TGF)-β1.
Methods The fibroblast cell line MRC-5 was seeded in a 96-well plate at a density of 5×103 cells/well, and stimulated by TGF-β1 at concentrations of 0, 2.5, 5.0, and 10.0 ng/mL for 24 h. The proliferation activity was measured by MTT method, and the relative mRNA levels of lncRNA-ATB, Col I, Col Ⅲ, and FN in stimulated MRC-5 cells were detected by real-time quantitative PCR. Spearman rank correlation test was used to assess the correlations of lncRNA-ATB mRNA levels with Col I, Col Ⅲ, and FN mRNA levels.
Results With the increasing of TGF-β1 dose, the proliferation activity of MRC-5 cells increased, and the cell proliferation activities of the 5.0 and 10.0 ng/mL groups were 1.07 and 1.12 times of the 0 ng/mL group (P < 0.05). With the increasing of TGF-β1 dose, the mRNA levels of FN in the 2.5, 5.0, and 10.0ng/mL groups were 2.11 (P=0.018), 3.58 (P=0.03), and 6.51 (P < 0.01) times of the 0ng/mL group, respectively; the mRNA levels of Col I in the 2.5, 5.0, and 10.0 ng/mL groups were 1.28 (P=0.324), 4.64 (P < 0.01), and 3.97 (P < 0.01) times of the 0 ng/mL group, respectively; the mRNA levels of Col Ⅲ in the 2.5, 5.0, and 10.0 ng/mL groups were 1.21 (P=0.400), 1.42 (P=0.120), and 4.23 (P < 0.01) times of the 0 ng/mL group, respectively; the mRNA levels of lncRNA-ATB in the 2.5, 5.0, 10.0 ng/mL groups were 1.80 (P=0.073), 3.57 (P < 0.01), and 4.55 (P < 0.01) times of the 0 ng/mL group, respectively. The results of correlation analysis showed that the mRNA level of lncRNA-ATB was positively correlated with the mRNA levels of FN (r=0.86), Col I (r=0.85) and Col Ⅲ (r=0.82) (Ps < 0.05).
Conclusion TGF-β1 could stimulate cell proliferation of fibroblasts, and increase the relative mRNA levels of lncRNA-ATB, Col I, Col Ⅲ, and FN. LncRNA-ATB is positively correlated with Col I, Col Ⅲ, and FN, suggesting that it may be related to the occurrence of pulmonary fibrosis.
DownLoad: