下调DKK2表达对小鼠子宫内膜基质细胞增殖和凋亡的影响
发布时间：2018-08-24 15:04
【摘要】：[目的]本试验旨在研究RNA干扰Dickkopf-2基因(DKK2)表达对小鼠子宫内膜基质细胞增殖和凋亡的影响。[方法]设计并合成3条针对DKK2基因的siRNA,在转染试剂LipofectamineTM2000介导下转染小鼠子宫内膜基质细胞,用RT-q PCR法检测转染前、后DKK2基因的表达量,筛选干扰效率最高的1条siRNA。用该siRNA转染小鼠子宫内膜基质细胞24 h,利用Caspase3活性和Annexin-V FITC/PI双染法检测干扰DKK2基因对细胞凋亡的影响;用CCK8细胞计数法检测该siRNA干扰DKK2基因不同时间对小鼠子宫内膜基质细胞增殖的影响。[结果]3条siRNA均能有效抑制DKK2基因的表达,RT-q PCR结果显示DKK2基因表达水平显著降低(siRNA2,P0.05;siRNA1、siRNA3,P0.01);CCK8法结果显示siRNA干扰DKK2后对小鼠子宫内膜基质细胞有明显的抑制作用(24 h、48 h,P0.05);Caspase3活性检测和Annexin-V FITC/PI双染法结果显示,siRNA1转染小鼠子宫内膜基质细胞24 h后,Caspase3活性明显增加(P0.01),细胞凋亡率明显增加(P0.01),凋亡率为18.48%。[结论]利用RNA干扰技术沉默DKK2基因的表达可以明显抑制小鼠子宫内膜基质细胞的增殖并促进凋亡。
[Abstract]:[objective] to study the effect of RNA interfering Dickkopf-2 gene (DKK2) expression on the proliferation and apoptosis of mouse endometrial stromal cells. [methods] three siRNA, targeting DKK2 gene were designed and synthesized to transfect mouse endometrial stromal cells mediated by transfection reagent LipofectamineTM2000. The expression of DKK2 gene before and after transfection was detected by RT-q PCR method, and one siRNA. with the highest interfering efficiency was screened. Mouse endometrial stromal cells were transfected with siRNA for 24 h. The effect of interfering DKK2 gene on apoptosis was detected by Caspase3 activity and Annexin-V FITC/PI double staining. The effects of siRNA interfering with DKK2 gene on the proliferation of mouse endometrial stromal cells were detected by CCK8 cell count method. [results] all three siRNA could effectively inhibit the expression of DKK2 gene RT-q PCR. The results showed that the expression level of DKK2 gene decreased significantly (siRNA2,P0.05;siRNA1,siRNA3,P0.01) CCK8 method showed that siRNA interference with DKK2 could inhibit the activity of Caspase-3 in mouse endometrial stromal cells (24 h, 48 h, P0.05). The results of detection and Annexin-V FITC/PI double staining showed that the activity of caspase3 and apoptosis of mouse endometrial stromal cells were significantly increased (P0.01), and the apoptotic rate was 18.48% after transfection of siRNA1 into mouse endometrial stromal cells for 24 h (P0.01). [conclusion] silencing the expression of DKK2 gene by RNA interference can significantly inhibit the proliferation of mouse endometrial stromal cells and promote apoptosis.
【作者单位】： 南京农业大学动物科技学院;
【分类号】：S814
，
本文编号：2201196
[Abstract]:[objective] to study the effect of RNA interfering Dickkopf-2 gene (DKK2) expression on the proliferation and apoptosis of mouse endometrial stromal cells. [methods] three siRNA, targeting DKK2 gene were designed and synthesized to transfect mouse endometrial stromal cells mediated by transfection reagent LipofectamineTM2000. The expression of DKK2 gene before and after transfection was detected by RT-q PCR method, and one siRNA. with the highest interfering efficiency was screened. Mouse endometrial stromal cells were transfected with siRNA for 24 h. The effect of interfering DKK2 gene on apoptosis was detected by Caspase3 activity and Annexin-V FITC/PI double staining. The effects of siRNA interfering with DKK2 gene on the proliferation of mouse endometrial stromal cells were detected by CCK8 cell count method. [results] all three siRNA could effectively inhibit the expression of DKK2 gene RT-q PCR. The results showed that the expression level of DKK2 gene decreased significantly (siRNA2,P0.05;siRNA1,siRNA3,P0.01) CCK8 method showed that siRNA interference with DKK2 could inhibit the activity of Caspase-3 in mouse endometrial stromal cells (24 h, 48 h, P0.05). The results of detection and Annexin-V FITC/PI double staining showed that the activity of caspase3 and apoptosis of mouse endometrial stromal cells were significantly increased (P0.01), and the apoptotic rate was 18.48% after transfection of siRNA1 into mouse endometrial stromal cells for 24 h (P0.01). [conclusion] silencing the expression of DKK2 gene by RNA interference can significantly inhibit the proliferation of mouse endometrial stromal cells and promote apoptosis.
【作者单位】： 南京农业大学动物科技学院;
【分类号】：S814
，
本文编号：2201196
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