Objective        To construct a lentivirus vector targeting fibroblast growth factor receptor 3 (FGFR3) gene and determine its effect on the proliferation and differentiation of prechondrogenic ATDC5 cells.        Methods        The lentivirus vector targeting FGFR3 gene was constructed and then trans

fected into 293T cells. The recombinant lentivirus vector was transfected into ATDC5 cells. The expression of FGFR3 was detected by real-time PCR and Western blotting. The effect of FGFR3RNAi vector on the proliferation of ATDC5 cells was measured by cell count assay and MTT assay. The effects of FGFR3RNAi vector on the makers of chondrogenic differentiation

including collagenⅡ

collagen Ⅹ and MMP-13

were detected by real-time PCR.        Results        The recombinant lentivirus vector targeting FGFR3 gene was successfully constructed. The lentivirus titer was 5×10

8

TU/mL. Real-time PCR showed that the FGFR3 mRNA level was decreased by 65.2% in FGFR3RNAi transfected cells compared with the cells transfected by the blank vector (P0.01)

and lower by 68.8% than the negative control (

P

0.01). Western blotting indicated that the expression of FGFR3 was also significantly reduced in FGFR3RNAi transfected cells than the blank vector-transfected cells and negative control (

P

0.01). Cell count assay and MTT assay demonstrated that FGFR3RNAi vector resulted in strongly inhibited proliferation in ATDC5 cells than the blank vector and negative control (

P

0.05

P

0.01). Real-time PCR indicated that FGFR3 induced the mRNA expression of collagenⅡ

collagenⅩ and MMP-13 than the other 2 kinds of cells (

P

0.01).        Conclusion       The recombinant lentivirus vector targeting FGFR3 gene is successfully constructed. It efficiently down-regulates FGFR3 in ATDC5 cells and alleviates the FGFR3-induced inhibition on the proliferation and differentiation in ATDC5 cells.