Silencing of Twist Expression by RNA Interference Suppresses Epithelial-mesenchymal Transition, Invasion, and Metastasis of Ovarian Cancer

Purpose: This study aimed to explore the role of the Twist gene in the epithelial-mesenchymal transition ofovarian cancer.
Methods: An RNA interference plasmid expressing a small interfering RNA (siRNA)-targetingTwist (Twist siRNA vector) was designed, constructed, and transfected into the human ovarian cancer cell lineA2780. Transfection efficiency was assessed under a fluorescence microscope. Changes in the expression of TwistmRNA in A2780 after transfection with the pGenesil Twist shRNA plasmid were analyzed through RT-PCR.MTT assays and adhesion experiments were applied to determine changes in proliferation and adhesion abilityof A2870 after transfection with the Twist shRNA plasmid. Changes in the expression of the E-cadherin andN-cadherin proteins in A2780 after transfection with the Twist shRNA plasmid were analyzed using Westernblotting.
Result: The restructuring plasmid pGenesil-Twist shRNA was constructed successfully. After 48 h ofculture, 80% of the cells expressed high-intensity GFP fluorescence and stability. The expression of Twist decreasedsignificantly after the transfection of the Twist shRNA plasmid (P<0.05). Proliferation of the transfected TwistshRNA cells showed no difference with that of the A2780-nontransfection or A2780-si-control groups (P>0.05) butthe adhesion ability of A2780 decreased dramatically (P<0.05). Expression of the E-cadherin protein increased,whereas that of the N-cadherin protein decreased compared with that in the A2780-nontransfection or A2780-si-control groups (P<0.05).
Conclusion: Twist is essential for epithelial-mesenchymal transition, invasion, andmetastasis of ovarian cancer.