Effect of interfering RNA targeting pituitary specific transcription factor-1 on biological behaviors in rat growth hormone pituitary adenoma cells

Abstract

Objective To observe the effect of small interfering RNA (siRNA) silencing pituitary specific transcription factor 1 (Pit-1) gene expression on the biological behaviors in the rat pituitary growth hormone adenoma cells (GH3).Methods Pit-l-targeted siRNA was designed and synthesized,and transfected into GH3 cells with mixed with liposome.Western blotting and reverse transcription polymerase chain reaction (RT-PCR) were applied to detect the expression levels of Pit-1 protein and mRNA after siRNA-Pit-1 transfection.Cell proliferation was examined by using cell counting kit-8 (CCK-8) cell proliferation-toxicity test.Cell apoptosis was detected by using flow cytometry.In vitro invasion ability changes in GH3 cells were detected by Transwell cell array.Results siRNA significantly inhibited the expression of Pit-1 protein and mRNA.As compared with negative control group,cell proliferation ability was decreased with the absorbance (A) values being respectively decreased by 0.46 and 0.41 at 36 and 48 h,apoptosis rate was increased by 3.17％,6.36％ and 8.46％,and the number of cells penetrating the membrane was decreased by 38.43,46.54 and 38.13 at 24,36 and 48 h in siRNA group,respectively (P ＜ 0.05 for all).Conclusion Pit-1 siRNA can effectively inhibit proliferation,promote apoptosis and reduce invasiveness of GH3 cells. Key words: Pituitary adenoma; Pituitary specific transcription factor-1