Effect Of Down-Regulating FNBP1 Gene On Cell Proliferation And Metastasis Ability In HL-7702 Cells

Objective: To construct the small interfering RNA ( siRNA) targeting FNBP1 with a high specificity and to study the effect of silencing FNBP1 gene on the proliferation and metastasis ability of HL-7702 cellsMethods: Three small interference expression vectors (Si-1,Si-2,Si-3) specific to FNBP1 mRNA were designed and constructed, then were transfected into the HL-7702 cells after identified by restriction analysis and DNA sequencing. Analysis of efficiency of FNBP1 siRNA was detected by RT-PCR and Western-blot to choose the highest inhibitory target of FNBP1 among the three vectors. The cell line with stable expression of siRNA of FNBP1 was selected by G418 medium for 15 days. After FNBP1 knockdown, the proliferation of HL-7702 cells was measured by the XTT assay and metastasis ability was measured by wound healing assay.Results: Restriction analysis and sequencing proved that siRNA expression vector constructed correctly; FNBP1 mRNA and protein expression obviously decreased in HL-7702 cells transfected with siRNA (Si-1,Si-2,Si-3) respectively compared with the control groups, and Si-2 have the highest inhibitory effect.Conclusion: XTT showed that Si-2 can significantly inhibit the proliferation in HL-7702 cells, and have no effect on metastasis ability of HL-7702, indicating that FNBP1 plays an important role in proliferation of HL-7702 cells.