过表达<em>NDRG1</em>对绵羊原代子宫内膜上皮细胞生物功能的影响

doi:10.3969/j.issn.1674-7968.2023.11.009

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摘要子宫内膜上皮细胞作为子宫内重要的细胞之一,其生物功能的改变影响着雌性动物妊娠的发生。为了探究N-myc下游调节基因1 (N-myc downstream regulated gene 1, NDRG1)的过表达对绵羊(Ovis aries)原代子宫内膜上皮细胞的生物功能的影响,本研究纯化原代绵羊子宫内膜上皮细胞,用瞬时转染的方法将NDRG1过表达载体导入原代子宫内膜上皮细胞,qRT-PCR和Western blot检测瞬时细胞模型的构建;用qRT-PCR检测细胞周期蛋白D1基因(cyclin D1, CCND1)、CCNB1和CCNA1、上皮间质转化(epithelial-mesenchymal transition, EMT)相关基因的表达;CCK-8法检测细胞的增殖能力,划痕实验检测细胞迁移能力。结果表明,pcDNA3.1-NDRG1组NDRG1基因及其蛋白的表达量极显著增高(P<0.01);CCND1表达量极显著下降(P<0.01);E-钙黏蛋白(E-cadherin)表达量极显著增高(P<0.01),波形蛋白基因(Vimentin)表达量极显著降低(P<0.01);CCK-8结果显示,pcDNA3.1-NDRG1组增殖能力被抑制;划痕实验结果显示,pcDNA3.1-NDRG1组抑制细胞迁移能力。综上表明,NDRG1的过表达抑制了细胞的增殖,阻断了细胞周期G1/S的转变,抑制了其迁移能力,提示这与EMT过程相关。本研究为促进绵羊早期妊娠成功提供研究材料。

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	龙德智
	郝科兴
	陈慧慧
	胡广东
	王静

关键词 ： N-myc下游调节基因1 (NDRG1), 原代子宫内膜上皮细胞, 绵羊

Abstract：Endometrial epithelial cells are important in the uterus, and their biological function changes affect the occurrence of pregnancy in female animals. To investigate the effect of over-expression of N-myc downstream regulatory gene 1 (NDRG1) on biological function of primary endometrial epithelial cells in sheep (Ovis aries), the primary of sheep endometrial epithelial cells were purificated and NDRG1 over-expression vector was transfected into primary endometrial epithelial cells by transient transfection, and the establishment of transient cell model was detected by qRT-PCR and Western blot. The expression of cyclin D1 (CCND1), CCNB1, and CCNA1 and epithelial-mesenchymal transition (EMT)-related genes was detected by qRT-PCR. CCK-8 assay was used to detect cell proliferation and scratch assay was used to detect cell migration. The purification effect of primary cells was good and could be used for subsequent experiments. The expression of NDRG1 and its protein in PCDNA3.1-NDRG1 group were extremely significantly increased (P<0.01). The expression of CCND1 decreased extremely significantly (P<0.01). The expression of E-cadherin was extremely significantly increased (P<0.01), and that of Vimentin was extremely significantly decreased (P<0.01); CCK-8 results showed that proliferation was inhibited in PCDNA3.1-NDRG1 group. The results of scratch treatment showed that pcDNA3.1-NDRG1 group inhibited cell migration. NDRG1 over-expression inhibited cell proliferation, blocked cell cycle G1/S transition, inhibited the process of EMT, and also inhibited cell migration ability to some extent, which provides a theoretical basis for studying the promotion of successful early pregnancy in sheep.

Key words： N-myc downstream regulated gene 1 (NDRG1) Primary endometrial epithelial cells Sheep

收稿日期: 2022-07-11

ZTFLH:

S826

基金资助:兵团重点领域科技攻关计划项目(2021AB014); 石河子大学创新发展专项(CXFZ202112)

通讯作者: *huguangdong1017@163.com;wjtry100@163.com

引用本文:

龙德智, 郝科兴, 陈慧慧, 胡广东, 王静. 过表达NDRG1对绵羊原代子宫内膜上皮细胞生物功能的影响[J]. 农业生物技术学报, 2023, 31(11): 2304-2312.
LONG De-Zhi, HAO Ke-Xing, CHEN Hui-Hui, HU Guang-Dong, WANG Jing. Effect of Overexpression of NDRG1 on Biological Function of Primary Endometrial Epithelial Cells in Sheep (Ovis aries). 农业生物技术学报, 2023, 31(11): 2304-2312.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2023.11.009 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2023/V31/I11/2304

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