Effects of Suppressor of Cytokine Signaling-1 Silencing on the Proliferation and Interferon Gamma Susceptibility of Human Pancreatic Cancer Cells / 天津医药

ABSTRACT

Objective To detect the changes of cell proliferation and IFN-γsusceptibility of human pancreatic can-cer cells after suppressor of cytokine signaling-1 (SOCS1) gene silencing, and to explore the SOCS1 as the target of anti-tu-mor therapy through enhancing the function of IFN-γ. Methods Western blot assay, PCR and real-time PCR were used to verify the down regulation of SOCS1 in human pancreatic cancer cell (PANC1) after transfection;subsequently, PANC1 was stimulated with IFN-γ. Western blot assay was also used to detect the expression of signal transducers and activators of tran-scription (STAT)1 and phosphorylation STAT(pSTAT)1;and the change of IFN-γsusceptibility was detected by MTT assay. Real-time PCR was used to detect the mRNA of interferon regulatory factor-1(IRF-1). Flow cytometry was used to detect the cell cycle. Results The expression levels of SOCS1 mRNA and protein were significantly decreased in small hairpin SCOS1 (shSOCS1) transfected PANC1 cells. After the silence of SOCS1, the expression levels of IRF-1 and pSTAT1 in-creased significantly (P<0.05), and the median inhibitory concentration(IC50)of IFN-γfor PANC1 cells decreased signifi-cantly (P<0.01). The cell count of shSOCS1 cells dropped significantly compared with that of control group after the SOCS1 silencing for 72 hours (P<0.05). The cell cycle arrest was promoted at the G0/G1 phase, but the percentage of cells in S phase and G2/M decreased compared to that of control groups (P<0.05). Conclusion After the inhibition of SOCS1 gene expression, the proliferation ability of human pancreatic cancer cell line PANC1 decreased, and the sensitivity of PANC1 cells to IFN-γwas enhanced.