Abstract: Objective: Based on neurotrophic domain sequence (NP) of saposin C, eukaryotic expression vectors containing NP sequence and NP fused to TAT transduction domain were respectively constructed. The TAT transmembrane signal was expected to help NP go through cell membranes and have effects on prostate cancer cells. Methods: The eukaryotic expression vectors pcDNA-TAT-NP and pcDNA-NP were constructed by using gene cloning technology. After transient transfection of pcDNA-TAT-NP and pcDNA-NP into the prostate cancer cells PC3 and LNCaP, RT-PCR, MTT and flow cytometry were used to respectively detect the expression of TAT-NP and NP in the level of mRNA, prostate cancer cell growth and the cell cycle changes. Results: The eukaryotic expression vectors pcDNA-TAT-NP and pcDNA-NP were constructed and confirmed by sequencing. The result obtained from RT-PCR indicated both pcDNA-TAT-NP and pcDNA-NP were expressed in prostate cancer cells. MTT analysis showed that expressions of TAT-NP and NP had a stimulating effect on prostate cancer cell proliferation, and promoted the cells to enter S and G₂/M phases. Conclusion: Ectopic expression of NP could enhance the prostate cancer cell growth and affect the cell cycle.

Key words: Prosaposin, Neurotrophic Peptide, TAT transduction domain, Flow cytometry, Prostate neoplasms, Cell lines