Objective: To explore the relationship between WWOX gene and attachment and adhesion of ovarian cancer.
Methods: The expression of WWOX mRNA was detected by RT-PCR, the expression of the WWOX protein was evaluated by western blot in WWOX-transfected PEO1 cells (H6, H7, H8 cell) and vector-transfected control cells (vec-1, vec-2 cell). Attachment assay was used to assess the adhesion of the transfection in PEO1 cells via culturing the cells on the pre-coated fibronectin wells. RNA interference (RNAi) was used to knockdown the endogenous expression of WWOX in the A2780 ovarian cancer cell line by liposome. Attachment assay was detected the adhesion to fibronectin after gene silencing.
Results: RT-PCR showed that expression of mRNA WWOX in exon9 was in all transfection cells (H6, H7, H8, vec-1, vec-2 cell). Western blot showed that expression of WWOX protein was in the WWOX-transfected cells (H6, H7, H8 cell), but not in the vector-transfected cells (vec-1, vec-2 cell). Attachment assay showed that H6, H7, H8 cell (0.098 +/-0. 003, 0.091 +/- 0.004, 0.099 +/- 0.003) adhered more slowly to fibronectin than vec-1, vec-2 cell (0.185 +/- 0.003, 0.175 +/- 0.006) and non-transfected PEO1 cell (0.211 +/- 0.007), and demonstrated significantly reduced adhesion after 2 hours (P < 0.01). A2780 adhesive cells that WWOX gene be knockdown was 0.059 +/- 0.005, adhered more significantly rapid than those untreated cells that was 0.029 +/- 0.003 after treated 30 minutes (P < 0.05).
Conclusions: WWOX gene can suppress adhesion to fibronectin in ovarian cancer cells. This suggests an important role for loss of WWOX gene in promoting attachment and adhesion of ovarian cancer cells on loco-regional peritoneum, and further resulting in enhancing loco-regional peritoneal tumor invasiveness and spread.