敲减STAT3基因表达对胰腺祖细胞增殖的影响

Effects of STAT3 knockdown on proliferation of pancreatic progenitor cells

目的:探讨敲减信号转导和转录激活因子3(STAT3)基因的表达对胰腺祖细胞增殖的影响。方法:将靶向STAT3基因的小干扰RNA(siRNA)转染胰腺祖细胞,采用qPCR和Western blot验证STAT3基因表达下调,通过光学显微镜、活细胞计数法和CCK8法检测STAT3基因沉默对胰腺祖细胞增殖和活力的影响,流式细胞术检测细胞周期进程的变化,Western blot检测细胞周期蛋白D2(CCND2)的表达水平。结果:干扰片段有效沉默STAT3基因的表达,STAT3的蛋白表达水平降低了(78.62±0.08)%,与对照组相比,敲减STAT3基因的表达使胰腺祖细胞数量减少了(45.62±0.03)%,活力降低了(43.68±0.05)%,细胞周期阻滞在G 0/G 1期,CCND2蛋白表达水平明显降低(P<0.05)。结论:敲减STAT3基因表达通过抑制CCND2蛋白表达,阻滞细胞周期在G0/G1期,抑制胰腺祖细胞的增殖和活力。

AIM:To investigate the effect of signal transducer and activator of transcription 3(STAT3)silencing on the proliferation of pancreatic progenitor cells.METHODS:The small intefering RNA(siRNA)targeting STAT3 gene was transfected into the pancreatic progenitor cells,and knock-down of STAT3 gene expression was verified by qPCR and Western blot.The effects of STAT3 silencing on the proliferation and viability of pancreatic progenitor cells were analyzed by optical microscopy,cell counting and CCK8 assay.The cell cycle distribution was detected by flow cytometry.The protein expression of cyclin D2(CCND2)was determined by Western blot.RESULTS:The interference fragment targeting STAT3 gene effectively silenced the expression of STAT3,and the protein expression level of STAT3 was decreased by(78.62±0.08)%.Compared with control group,knock-down of STAT3 expression decreased the number of pancreatic progenitor cells by(45.62±0.03)%,reduced the viability of the cells by(43.68±0.05)%,arrested the cell cycle at G0/G1 stage,and decreased the protein expression of CCND2.CONCLUSION:Knock-down of STAT3 expression arrests the cell cycle at G0/G1 stage and inhibits the proliferation and viability of pancreatic progenitor cells by inhibiting the protein expression of CCND2.