Objective: To assess the suppression effect on WWOX gene in SKOV3/SB cell line by small interference RNA (siRNA).
Methods: Transfection of siRNA using lipofectamine 2000 was conducted to silence WWOX gene expression, the expression levels of WWOX mRNA and protein were evaluated, and the effects on the cell cycles at 48 hours of transfection were assessed by RT-PCR, western blot and flow cytometry (FCM) respectively. The cisplatin resistance index was assayed after transfection of SKOV3/SB with siRNA by methyl thiazolyl tetrazolium (MTT) and the cisplatin concentration of SKOV3/SB cells transfected with siRNA of WWOX was measured by high performance liquid chromatography.
Results: (1) In SKOV3/SB cells transfected with WWOX interference fragment, whether at the mRNA or protein level, the expression of both of WWOX decreased. There was a significant difference (P < 0.05) compared with SKOV3 cells and non-transfected cells. (2) After transfection of the WWOX interference fragment, the index of platinum resistance of SKOV3/SB decreased from 5.04 to 3.89. (3) The number of cell transfected with the WWOX interference fragment in G(1) phase was increased, while that in S-phase was decreased. (4) The cisplatin concentration of SKOV3/SB cells transfected with the WWOX interference fragment was increased from 9.43 ng/L to 23.45 ng/L compared with SKOV3/SB cells non-transfected with a significant difference (P < 0.05).
Conclusion: WWOX gene may be involved in cisplatin resistance phenomenon in epithelial ovarian cancer.