Effects of SOCS1 gene overexpression on proliferation and apoptosis of porcine mammary epithelial cells
[Objective]The purpose of the study was to clarify the effect of overexpressed cytokine signal transducer suppressor 1(SOCS1)gene on the expression of key signaling pathway(JAK/STAT)related genes in mammary gland de-velopment,and to provide reference for revealing the mechanism of SOCS1 gene regulating proliferation and apoptosis of porcine mammary epithelial cells.[Method]The overexpressed vector pcDNA3.1-SOCS1 and the negative control vector pcDNA3.1-NC were constructed and transfected into porcine mammary epithelial cells,respectively.Then,cell prolifera-tion was detected by CCK-8,cell cycle and apoptosis were detected by flow cytometry,and the expression changes of JAK/STAT signaling pathway related genes after transfection were detected by real-time fluorescence quantitative PCR.[Result]Porcine mammary epithelial cells were transfected with overexpressed vector pcDNA3.1-SOCS1 and negative control vector pcDNA3.1-NC,respectively.The relative expression of SOCS1 gene in the overexpressed vector pcDNA3.1-SOCS1 transfection group was extremely significantly higher than that in the negative control vector pcDNA3.1-NC transfection group(P＜0.01,the same below).After transfection for 48 and 72 h,the OD value of porcine mammary epithelial cells in the overexpressed vector pcDNA3.1-SOCS1 transfection group was extremely significantly lower than that in the negative control vector pcDNA3.1-NC transfection group,indicating that the overexpressed vector pcDNA3.1-SOCS1 could effectively inhibit the proliferation ability of porcine mammary epithelial cells.The apoptosis rate of cells in the overexpressed vector pcDNA3.1-SOCS1 transfection group[(36.15±0.92)％]was significantly higher than that of the negative control vector pcDNA3.1-NC transfection group[(30.55±0.35)％](P＜0.05,the same below),and the rate of G1 phase cells in the overexpressed vector pcDNA3.1-SSOCSS1 transfection group was extremely signifi-cantly decreased,while the rates of S and G2 phase cells were significantly increased.Compared with the negative control vector pcDNA3.1-NC transfection group,except for JAK2,STAT3 and TYK2 genes,the relative expressions of 9 JAK/STAT signaling pathway related genes,including SOCS2,SOCS3,JAK1,STAT2,STAT5A,IL-6,PRLR,MCL1 and PIAS3,in the overexpressed vector pcDNA3.1-SSOCS1 transfection group were extremely significantly increased.[Con-clusion]Overexpression of SOCS1 gene can inhibit the proliferation of porcine mammary epithelial cells and promote the apoptosis of cells and affect the expression changes of JAK/STAT signaling pathway related genes,thus playing an impor-tant role in the regulation mechanism of mammary gland development in pigs.
万方数据
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