Abstract: Objective To build the lentiviral expression vector of pLV-EGFP (2A) puro-TrkB (Tyrosine kinase receptor B), establish the renal cell carcinoma cell line with stable overexpression of TrkB and observe the effect of TrkB on the proliferation of 786-O. Methods Coding sequence (CDS) of TrkB was amplified and combined with pLV-EGFP (2A) puro, then the connected product was transfected into human embryonic kidney HEK293V cell.Western blotting was used to detect TrkB protein level.The effect of TrkB on the proliferation of 786-O was detected by MTS assay and clone formation assay. Results pLV-EGFP(2A) puro-TrkB lentiviral expression vector was successfully constructed and overexpression of renal cell carcinoma cell line 786-O was achieved after transfecting with target cells.The expression of TrkB significantly up-regulated protein level compared with empty vector group.MTS assay showed that the absorbance in recombinant vector group decreased significantly after transfection (P< 0.05) and cell clone formation assay showed that cell colony numbers in recombinant vector group also decreased significantly compared with the empty vector group (P< 0.05). Conclusion The overexpression of TrkB may significantly decrease cell proliferation of 786-O cell line.