猪PLAG1基因过表达对3T3-L1前脂肪细胞增殖分化的影响  

Effects of Overexpression of Porcine(Sus Scrofa) PLAG1 Gene on Proliferation and Differentiation of 3T3-L1 Preadipocytes

作　　者：吴姿仪 白利瑶 吴嘉浩 窦腾飞 李新建[1] 韩雪蕾[1] 乔瑞敏[1] 王克君 杨峰[1] 李秀领 WU Zi-Yi;BAI Li-Yao;WU Jia-Hao;DOU Teng-Fei;LI Xin-Jian;HAN Xue-Lei;QIAO Rui-Min;WANG Ke-Jun;YANG Feng;LI Xiu-Ling(College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物科技学院,郑州450046

出　　处：《农业生物技术学报》2025年第4期782-791,共10页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划(2021YFD1301201);河南高等学校重点科研项目(23A230012);河南省农业良种联合攻关项目(2022020101)。

摘　　要：多形性腺瘤基因1 (plemorphic adenoma gene 1,PLAG1)在调控畜禽生长发育中具有重要作用。本课题组前期通过全基因组关联分析(genome-wide association study,GWAS)方法筛选到该基因与猪(Sus scrofa)的背膘厚(backfat thickness,BF)有关,其可能对脂肪细胞有潜在的调控作用。为进一步探究PLAG1基因与脂肪细胞的关系,本实验以3T3-L1前脂肪细胞为研究对象,并对PLAG1基因进行保守性分析,通过qRT-PCR、CCK-8、流式细胞术和油红O染色等技术探究过表达PLAG1基因对脂肪细胞增殖分化的影响。结果表明,PLAG1基因的保守性较高,在不同物种间的相似度为80%~92%;qRT-PCR结果显示,过表达PLAG1基因后,3T3-L1细胞增殖相关基因的表达极显著高于对照组(P<0.01);CCK-8结果表明过表达PLAG1基因后极显著提高了细胞活性(P<0.01);流式细胞术发现过表达PLAG1基因后S期的细胞数显著增加(P<0.05)。油红O染色结果显示,过表达PLAG1基因后显著降低成脂分化相关基因的表达(P<0.05),细胞脂滴数量也极显著降低(P<0.01)。综上,过表达PLAG1基因促进3T3-L1前脂肪细胞增殖,抑制其分化,说明PLAG1基因可能在脂肪细胞增殖和分化过程中发挥作用。本研究为进一步探究猪PLAG1基因调控脂肪沉积分子机制提供理论参考。Pleomorphic adenoma gene 1(PLAG1) plays a crucial role in regulating the growth and development of livestock and poultry.Our previous study identified that PLAG1 was associated with backfat thickness(BF) in pigs by genome-wide association study(GWAS),suggesting a potential regulatory effect on adipocytes.To further investigate the relationship between PLAG1 gene and adipocytes,3T3-L1 cells was used as a research model and PLAG1 conservation was analyzed.The effects of PLAG1 gene overexpression on cell proliferation and differentiation were examined using qRT-PCR,CCK-8 assays,flow cytometry,and Oil Red O staining.Results revealed high conservation of the PLAG1 gene,with sequence similarity among different species ranging from 80% to 92%.qRT-PCR analysis showed that,compared with the control group,the expression of proliferation-related genes in 3T3-L1 cells was extremely significantly increased following PLAG1 overexpression(P<0.01).CCK-8 assays showed that PLAG1 overexpression extremely significantly enhanced cell viability(P<0.01).Flow cytometry demonstrated a significant increase in the number of cells in the S phase following PLAG1 overexpression(P<0.05).Furthermore,Oil Red O staining indicated that the expression of adipogenic differentiation-related genes was significantly decreased after PLAG1 overexpression(P<0.05),accompanied by a extremely significant reduction in lipid droplet formation(P<0.01).In summary,PLAG1 overexpression promoted the proliferation of 3T3-L1 cells while inhibiting their differentiation,indicating that PLAG1 gene might play a regulatory role in the proliferation and differentiation of adipocytes.This study provides a theoretical foundation for further investigation of the molecular mechanisms through which the PLAG1 gene regulates fat deposition in pigs.

关 键 词：多形性腺瘤基因1(PLAG1) 猪 增殖分化 基因功能 3T3-L1前脂肪细胞 

分 类 号：S828.99[农业科学—畜牧学]