Effects Of Small Interfering RNA On CXCR4 Expression In LNCaP Cells

Objective: To study the effects of small interfering RNA(siRNA) on human CXCR4 expression in prostate cancer cell line (LNCaP cell). Methods: A potential target sequence of CXCR4 gene was selected for the synthesis of DNA templates in vitro. The two DNA stands were annealed into dsRNA, and then, the dsDNA was cloned into Psilencer 2.1-U6 neo vector. The recombinant plasmid Psilencer-siCXCR4 was transfected into LNCaP cells by lipofectamine 2000. The cells was collected after 48h. And the cellular CXCR4 expression was detected by RT- PCR. Two controls was seted up. One was transfected with no plasmid. And the other was transfected with negative plasmid Psilencer-C. Results: Restricion endonuclease digestion and DNA sequencing showed that siRNA plasmids based on Psilencer 2.1-U6 neo was constructed successfully. The siRNA could obviously suppress the expression of CXCR4. When transfected with plasmid Psilencer-siCXCR4 for 48 h, the levels of the expression of CXCR4 were reduced by(75.00±3.16)% respectively (P<0.01). The relative levels are as follows: 0.61±0.03 (transfected with nothing); 0.59±0.04 ( negative control);0.15±0.01 (transfected with Psilencer-siCXCR4). Conclusions: The siRNA express vector could successfully inhibit the expression of CXCR4 in prostate cancer cells . And it can be potentially useful in the next research of SDF-1/CXCR4 and prostate cancer.