小干扰质粒干扰肌腱膜纤维肉瘤癌基因同源物A对小鼠胰岛β细胞增殖及胰岛素基因表达的影响

发布时间：2018-05-03 10:33

本文选题：MIN细胞 + 肌腱膜纤维肉瘤癌基因同源物A　；参考：《中国糖尿病杂志》2017年06期

【摘要】：目的探讨肌腱膜纤维肉瘤癌基因同源物A(MafA)对胰岛β细胞增殖及胰岛素基因表达的影响。方法采用RT-PCR和Western blot分别测Ma。fA mRNA和蛋白的表达,筛选最佳小干扰质粒(siRNA),分为转染对照组(Cy3组)、阴性对照组(Scramble组)、阳性对照组(siβ-actin组)及实验组(即siMafA-1、siMafA-2和siMafA-3组)。按不同葡萄糖浓度分为5.6 mmol/L、10 mmol/L、25 mmol/L。根据不同葡萄糖浓度是否加入siRNA分为未加siRNA的对照组:A_0组5.6 mmol/L、B_0组10 mmol/L、C_0组25 mmol/L;加入siRNA的实验组:A_1组5.6 mmol/L、B_1组10 mmol/L、C_1组25 mmol/L。测MafA基因沉默后MIN6细胞增殖、胰岛素(Insulin)-1和Insulin-2 mRNA表达、MafA蛋白表达及上清液胰岛素浓度的变化。结果siMafA-2组干扰效果最好。C_1组MafA蛋白表达较C_0组下降(P0.05);MIN6细胞增殖、Insulin-2 mRNA表达和上清液胰岛素浓度均降低(P0.05)。A_1组与A_0组比较,MafA蛋白表达比较,差异无统计学意义(P0.05);MIN6细胞增殖、Insulin-2 mRNA表达和上清液胰岛素浓度比较,差异无统计学意义(P0.05)。不同糖浓度各亚组与对照组Insulin-1 mRNA表达比较,差异均无统计学意义(P0.05)。结论siRNA-2组是理想的siRNA。MafA下调可抑制小鼠胰岛β细胞增殖。MafA下调可抑制小鼠Insulin2 mRNA表达和胰岛素分泌。
[Abstract]:Objective to investigate the effects of oncogene congener Agna MafA on the proliferation and insulin gene expression of islet 尾 cells in tendon fibrosarcoma. Methods RT-PCR and Western blot were used to detect the expression of Ma.fA mRNA and protein respectively, and the best small interference plasmid siRNAs were screened. They were divided into three groups: transfection control group (Cy3), negative control group (scramble group), positive control group (sisi 尾 -actin group) and experimental group (siMafA-1 siMafA-2 and siMafA-3 group). There are 5.6 mmol / L 10 mmol 路L ~ (-1) 10 mmol / L ~ (10) mmol 路L ~ (-1) and 25 mmol / L ~ (-1) glucose concentration. According to whether different glucose concentrations were added to siRNA, they were divided into 5. 6 mmol / L / L group without siRNA, 5. 6 mmol / L / L\ + 1 group without siRNA and 10 mmol / L / L\% C\ + 0\ + 10 mmol / L\% C\% The proliferation of MIN6 cells, the expression of MafA protein and the concentration of insulin in supernatant were measured after MafA gene silencing. Results the interference effect of siMafA-2 group was the best. The expression of MafA protein in CST1 group was lower than that in CSTO group. The expression of insulin-2 mRNA and insulin concentration in supernatant of CST1 group were lower than that of CSTO group, and the expression of MafA protein in group A was lower than that in group A 0. There was no significant difference in the expression of insulin-2 mRNA and the insulin concentration in supernatant. There was no significant difference in the expression of insulin-2 mRNA and the concentration of insulin in supernatant. There was no significant difference in the expression of Insulin-1 mRNA between the subgroups with different glucose concentrations and the control group (P 0.05). Conclusion the down-regulation of siRNA.MafA in siRNA-2 group can inhibit the proliferation of islet 尾 cells in mice. MafA down-regulation can inhibit the expression of Insulin2 mRNA and insulin secretion in mice.
【作者单位】：福建医科大学附属漳州市医院内分泌科;湖南医药学院第一附属医院内分泌科;福建医科大学附属漳州市医院病理科;
【基金】：漳州市自然科学基金(ZZ2013J31)
【分类号】：R587.1

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