Construction of bovine TXNIP overexpression vector and its effect on proliferation and apoptosis of mammary epithelial cells
10.16303/j.cnki.1005-4545.2024.12.16
- VernacularTitle:牛TXNIP基因的过表达载体构建及对乳腺上皮细胞增殖和凋亡的影响
- Author:
Xiaoting WU
1
;
Xianji XU
1
;
Zonggang LUO
1
;
Tingting SUN
1
;
Ling WANG
1
Author Information
1. 西南大学动物科学技术学院,重庆荣昌 402460
- Publication Type:Journal Article
- Keywords:
TXNIP gene;
mammary epithelial cells;
cell proliferation;
apoptosis;
bovine
- From:
Chinese Journal of Veterinary Science
2024;44(12):2612-2618
- CountryChina
- Language:Chinese
-
Abstract:
To investigate the regulatory effect of thioredoxin-interacting protein(TXNIP)on the proliferation and apoptosis of bovine mammary epithelial cells,a bovine TXNIP gene overexpres-sion vector was constructed using the homologous recombination method.The overexpressed re-combinant plasmid was verified through enzyme digestion and sequencing,and the obtained se-quenccs were subjected to BLAST analysis and phylogenetic construction.The TXNIP overcxpres-sion plasmid was then transfected into bovine mammary epithelial cells.Cell proliferation was as-sessed using CCK-8 and EdU assays,while apoptosis was evaluated using Annexin V-mCherry.Ad-ditionally,RT-qPCR was employed to measure changes in the mRNA expression of apoptosis-re-lated genes in bovine mammary epithelial cells.The results showed that the CDS region of bovine TXNIP gene was 1 176 bp,encoded 391 amino acids,which was closely related to sheep.Compared to the control group,overexpression of the TXNIP gene decreased the viability of mammary epi-thelial cells,reduced cell proliferation,and promoted apoptosis.Specifically,overexpression of the TXNIP gene led to a significant upregulation of apoptosis-related genes,including Caspasc-3,Caspase-9,and Bax,in bovine mammary epithelial cells(P＜0.05).In contrast,the expression of Caspase-8 had no significant changes(P＞0.05).Additionally,the expression of anti-apoptosis gene Bcl-2 was significantly decreased(P＜0.01),and the ratio of Bax/Bcl-2 was significantly increased(P＜0.01).In conclusion,the TXNIP gene can inhibit the proliferation of bovine mam-mary epithelial cells,promote apoptosis,and regulate the expression of apoptosis-related genes,thereby influencing the development of the bovine mammary gland.
