Effects of Wnt3a on the CORM-3-induced osteogenic
differentiation of rat BMSCs
A successful transfection with pcDNA3.1-Wnt3a or Wnt3a siRNA was
confirmed by Western blotting (Fig. 5A) analysis. With overexpression of
Wnt3a, the mRNA levels of Runx2 and OPN were increased by 1.34- and
1.82-fold respectively on the 3rd day (P < 0.05), and
1.40- and 1.29-fold respectively on the 7th day of osteogenic induction,
compared with CORM-3 group (P < 0.05) (Fig. 5B). On the
contrary, in Wnt3a-deficient cells, mRNA expressions of Runx2 and OPN
were all reduced on the 3rd and the 7th day of osteogenic induction
(P < 0.05) (Fig. 5C). The protein expressions of Runx2
and OPN after the osteogenic induction were similarly regulated by
Wnt3a, in line with the regulation of Wnt3a on the mRNA expression (Fig.
5D-E). In Wnt3a-overexpression cells, the protein expressions of Runx2
and OPN were increased by 1.60- and 1.85-fold respectively on the 3rd
day, 2.02- and 2.56-fold respectively on the 7th day of osteogenic
induction, compared with CORM-3 group (P < 0.05).
Moreover, mineralization in Wnt3a-overexpression cells on the 14th day
of osteogenic induction was enhanced significantly, compared with CORM-3
group. The opposite effect was observed in Wnt3a-deficient cells
(P < 0.05) (Fig. 5F).