【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。更多还原

【Abstract】 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.更多还原