Abstract:Objective:To inhibit growth hormone receptor (GHR) gene expression in human colon cell line SW480 by small interfering RNA (siRNA) technique, and thereby to observe the effect of GHR gene silencing on proliferation of SW480 cells.
Methods:After the GHR gene specific siRNA design, the eukaryotic plasmid expression vector targeting human GHR (pcDNATM6.2-GW/EmGFP-siRNA-GHR) was constructed, which was then transfected into SW480 cells by electroporation. The transfected cells were cultured for 48 to 72 hours, and then the cell proliferative activity was assayed by tetrazolium bromide (MTT), GHR protein expression was detected by Western Blot, and GHR mRNA was determined by fluorescence real time quantitative reverse transcription-polymerase chain reaction, respectively.
Results:Compared with the control group, the proliferative speed, GHR protein and mRNA expression of SW480 cells were all significantly decreased after transfection (all P<0.05).
Conclusions: The pcDNATM6.2-GW/EmGFP-GHR-siRNA constructed can effectively down-regulate GHR expression in SW480 cells, and thereby inhibit the proliferation of SW480 cells.