Overexpression of EBP50 Suppresses the proliferation of FLS
Inhibiting abnormal proliferation and activation of FLS may be a
potentially therapeutic strategy to block or delay the progression of
RA. To investigate the potential
effects of EBP50 on the proliferation of FLS, fls were
transfected with LV-EBP50 or LV-Plvx
for 72 h, then analyzed for the EBP50 expression and the proliferation
of FLS. The results showed that LV-EBP50 transfection significantly
increased the expression of EBP50 in both mRNA levels (Fig. 1A) and
protein levels (Fig. 1B-1C), and MTT assay demonstrated that
overexpression of EBP50 can suppresse the proliferation of FLS
significantly, when compared with untransfected cells and LV-Plvx
transfected cells (Fig. 1D). It has been reported that EBP50 can inhibit
the proliferation of breast cancer cells by stimulating the autophagic
lysosomal degradation, so we speculate whether EBP50 inhibits FLS cell
proliferation by inhibiting autophagy. Then, the expression of LC3 was
measured by western blotting in LV-EBP50 and LV-plvx transfected cells
and untreated cells.As shown in (Fig. 1E-1F), the expressions of LC3 in
FLS remain unchanged after transfected with LV-EBP50.Collectively, these
results suggest that overexpression of EBP50 suppresses the
proliferation of FLS and has no influence on autophagy.
Overexpression of EBP50
promote apotosis of FLS By activating caspase-3
FLS can exhibit tumor-like behavior, such as significant inhibition of
apoptosis. As the major tumor suppressor genes, related literature
reports have indicated that overexpression of EBP50 promoted
significantly the tumor cells apotosis, and our previous results have
showed that the overexpression of EBP50 upregulates the apoptosis ratio
of macrophage by increasing the expression of Bax and caspase-3 in an
NO-dependent mechanism. To investigated whether EBP50 can perform the
same function in FLS, FLS were transfected with LV-EBP50 and LV-Plvx at
the MOIs of 10 for 48 h, 72 h, 96 h, and the cell apoptosis of FLS was
analyzed by FCM. Results showed that EBP50 overexpression increased
distinctly the apoptosis of FLS when compared with LV-Plvx transduced
FLS (P< 0.001) (Fig. 2A); The expression of apoptosis-related
protein caspase-3 in FLS were measured by western blotting. As shown in
(Fig. 2B), when FLS were transduced with LV-EBP50 and LV-Plvx for 96h
,the protein expressions level of cleaved caspase-3 were significantly
increased .
overexpression of EBP50Suppresses
the Migration and Invasion
of FLS