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Effect of Overexpression of NDRG1 on Biological Function of Primary Endometrial Epithelial Cells in Sheep (Ovis aries) |
LONG De-Zhi, HAO Ke-Xing, CHEN Hui-Hui, HU Guang-Dong*, WANG Jing* |
College of Animal Science and Technology, Shihezi University, Shihezi 832000, China |
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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.
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Received: 11 July 2022
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Corresponding Authors:
* Corresponding authors, huguangdong1017@163.com; wjtry100@163.com
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