农业生物技术学报2024，Vol.32Issue(10)：2293-2305,13.DOI:10.3969/j.issn.1674-7968.2024.10.009
LOC112447378/miR-15a/PRLR信号轴对MAC-T细胞增殖及凋亡的调控及机制
Regulation and Mechanism of LOC112447378/miR-15a/PRLR Signaling Axis on Proliferation and Apoptosis of MAC-T Cells
摘要
Abstract
Global warming has made the issue of heat stress in cows(Bos taurus)increasingly prominent,causing huge economic losses to the dairy industry.Elucidating the molecular mechanisms underlying the occurrence and response of heat stress in cows can better address the issue of heat stress.Differentially expressed long non-coding RNA(lncRNA),microRNA(miRNA),and mRNA profiles between heat and non-heat stressed conditions were screened using high-throughput sequencing in our previous study.In this study,LOC112447378/miR-15a/PRLR was identified as a key competing endogenous RNA(ceRNA)network in heat stress response through bioinformatics analysis based on the high-throughput data.By constructing an in vitro heat stress model using mammary alveolar cells-large T antigen(MAC-T),the mRNA expression levels of LOC112447378,miR-15a,and prolactin receptor(PRLR)were determined using qRT-PCR.The protein levels of PRLR were determined using Western blot.These results were consistent with high-throughput sequencing results.The mRNA expression level of LOC112447378 was significantly downregulated when miR-15a was overexpressed;The mRNA and protein levels of PRLR were significantly downregulated when miR-15a was overexpressed.After knocking down miR-15a,the mRNA expression level of LOC112447378 was significantly upregulated;The mRNA and protein levels of PRLR were significantly upregulated.The methyl thiazolyl tetrazolium(MTT)test results showed that miR-15a inhibited the proliferation of MAC-T cells.The results of Annexin V-FITC/PI dual staining showed that miR-15a promotes apoptosis of MAC-T cells.MiR-15a could target and regulate the target genes LOC112447378 and PRLR,thereby achieving regulatory effects of inhibiting MAC-T cell proliferation and promoting apoptosis.These results indicated that the LOC112447378/miR-15a/PRLR signaling axis could affect the proliferation and apoptosis of MAC-T cells,thereby playing an important regulatory role in the heat stress response process of MAC-T cells.This study provides new ideas and theoretical basis for molecular breeding of heat-resistant and high-yield cows in the future.关键词
热应激/牛乳腺上皮细胞(MAC-T)/miR-15a/催乳素受体(PRLR)/长链非编码RNA(lncRNA)Key words
Heat stress/Mammary alveolar cells-large T antigen cells(MAC-T)/miR-15a/Prolactin receptor(PRLR)/Long non-coding RNA(lncRNA)分类
农业科技引用本文复制引用
高梦静,韩承芮,张晓雨,郭月美,刘思萌,谢圣浩,李秋玲,齐颖..LOC112447378/miR-15a/PRLR信号轴对MAC-T细胞增殖及凋亡的调控及机制[J].农业生物技术学报,2024,32(10):2293-2305,13.基金项目
河北省自然科学基金青年科学基金(C2024408013) （C2024408013）
河北省高等学校科学技术研究重点项目(ZD2021091) （ZD2021091）
廊坊师范学院优秀科研创新团队(JYT202301) （JYT202301）
廊坊师范学院优秀青年项目(JYQ202301) （JYQ202301）
大学生创新创业训练计划项目(X202310100014) （X202310100014）
