畜牧与兽医2025，Vol.57Issue(5)：1-8,8.
干扰OPN基因对贵州黑山羊卵巢颗粒细胞增殖、凋亡及繁殖相关基因的影响
Effects of interference with OPN gene on proliferation,apoptosis and reproduction related genes of ovarian granulosa cells in Guizhou black goat
摘要
Abstract
The aim of this study was to investigate the relationship between the osteopontin(OPN)gene and proliferation and apoptosis of ovarian granulosa cells(OGCs)in Guizhou black goats.Using Guizhou black goat as the research object,we screened the interfering vectors by constructing four interfering vectors of OPN gene,transfecting them into OGCs,and detecting the inhibition efficiency of different interfer-ing vectors on the OPN gene by real-time fluorescence quantitative PCR(RT-qPCR).We used cell counting kit-8(CCK8)to detect the proliferation viability of the cells,and the cell scratch assay to detect the migration ability.And we detected the expression of the reproductive marker genes and the apoptosis-related genes associated with them by fluorescence quantitative PCR.In this study,four interference vectors of the OPN gene were successfully constructed.shRNA-OPN-222,shRNA-OPN-385 and shRNA-OPN-741 all suppressed the gene ex-pression of the OPN extremely significantly,compared with the shRNA-NC group(P＜0.01),and the suppression effect of shRNA-OPN-222 was the most effective.The results of the CCK8 and cell scratch assay showed that shRNA-OPN-222 highly significantly inhibited the proliferative vigour and migration ability of OGCs,compared with the shRNA-NC group(P＜0.01).The RT-qPCR results showed that si-lencing the OPN gene highly significantly inhibited the gene expression of the DCN2,CYP19A1 and ESR1(P＜0.01),significantly promo-ted the gene expression of CTNNB2,Caspase-3 and Bax(P＜0.05),and had no significant effect on the gene expression of the FTH1,Caspase-9.In conclusion,this study indicated that interference with the OPN gene effectively inhibited the proliferation and migration of OGCs,lowered the expression level of the reproduction-related genes,and simultaneously promoted the apoptotic process of the cells.These results laid a foundation for further elucidation of the molecular mechanism by which the OPN gene affects the reproductive rate of goats.关键词
OPN基因/贵州黑山羊/卵巢颗粒细胞/细胞增殖/繁殖相关基因Key words
OPN gene/Guizhou black goat/ovarian granulosa cell/cell proliferation/reproduction-related genes分类
畜牧业引用本文复制引用
马丝,夏薇,温晓艳,潘智仁,付业海,刘彬,陆情梅,赵佳福..干扰OPN基因对贵州黑山羊卵巢颗粒细胞增殖、凋亡及繁殖相关基因的影响[J].畜牧与兽医,2025,57(5):1-8,8.基金项目
大学生SRT计划项目(贵大SRT字[2022]085号) （贵大SRT字[2022]085号）
贵州省科技计划项目(黔科合支撑[2022]一般089) （黔科合支撑[2022]一般089）
贵州省"生物育种先导性研究"项目(黔科合支撑[2022]重点033) （黔科合支撑[2022]重点033）
贵州省科技创新团队项目(黔科合平台人才-CXTD[2023]025) （黔科合平台人才-CXTD[2023]025）
