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Abstract：
BACKGROUND: Polycystic ovary syndrome (PCOS) is one of the most complex endocrine disorders in women of reproductive age. Abnormal proliferation of granulosa cells (GCs) is an important cause of PCOS. This study aimed to explore the role of fatty acid-binding protein 5 (FABP5) in granulosa cell (GC) proliferation in polycystic ovary syndrome (PCOS) patients.
METHODS: The FABP5 gene, which is related to lipid metabolism, was identified through data analysis of the gene expression profiles of GSE138518 from the Gene Expression Omnibus (GEO) database. The expression levels of FABP5 were measured by quantitative real-time PCR (qRT‒PCR) and western blotting. Cell proliferation was evaluated with a cell counting kit-8 (CCK-8) assay. Western blotting was used to assess the expression of the proliferation marker PCNA, and immunofluorescence microscopy was used to detect Ki67 expression. Moreover, lipid droplet formation was detected with Nile red staining, and qRT‒PCR was used to analyze fatty acid storage-related gene expression.
RESULTS: We found that FABP5 was upregulated in ovarian GCs obtained from PCOS patients and PCOS mice. FABP5 knockdown suppressed lipid droplet formation and proliferation in a human granulosa-like tumor cell line (KGN), whereas FABP5 overexpression significantly enhanced lipid droplet formation and KGN cell proliferation. Moreover, we determined that FABP5 knockdown inhibited PI3K-AKT signaling by suppressing AKT phosphorylation and that FABP5 overexpression activated PI3K-AKT signaling by facilitating AKT phosphorylation. Finally, we used the PI3K-AKT signaling pathway inhibitor LY294002 and found that the facilitation of KGN cell proliferation and lipid droplet formation induced by FABP5 overexpression was inhibited. In contrast, the PI3K-AKT signaling pathway agonist SC79 significantly rescued the suppression of KGN cell proliferation and lipid droplet formation caused by FABP5 knockdown.
CONCLUSIONS: FABP5 promotes active fatty acid synthesis and excessive proliferation of GCs by activating PI3K-AKT signaling, suggesting that abnormally high expression of FABP5 in GCs may be a novel biomarker or a research target for PCOS treatment.
METHODS: The FABP5 gene, which is related to lipid metabolism, was identified through data analysis of the gene expression profiles of GSE138518 from the Gene Expression Omnibus (GEO) database. The expression levels of FABP5 were measured by quantitative real-time PCR (qRT‒PCR) and western blotting. Cell proliferation was evaluated with a cell counting kit-8 (CCK-8) assay. Western blotting was used to assess the expression of the proliferation marker PCNA, and immunofluorescence microscopy was used to detect Ki67 expression. Moreover, lipid droplet formation was detected with Nile red staining, and qRT‒PCR was used to analyze fatty acid storage-related gene expression.
RESULTS: We found that FABP5 was upregulated in ovarian GCs obtained from PCOS patients and PCOS mice. FABP5 knockdown suppressed lipid droplet formation and proliferation in a human granulosa-like tumor cell line (KGN), whereas FABP5 overexpression significantly enhanced lipid droplet formation and KGN cell proliferation. Moreover, we determined that FABP5 knockdown inhibited PI3K-AKT signaling by suppressing AKT phosphorylation and that FABP5 overexpression activated PI3K-AKT signaling by facilitating AKT phosphorylation. Finally, we used the PI3K-AKT signaling pathway inhibitor LY294002 and found that the facilitation of KGN cell proliferation and lipid droplet formation induced by FABP5 overexpression was inhibited. In contrast, the PI3K-AKT signaling pathway agonist SC79 significantly rescued the suppression of KGN cell proliferation and lipid droplet formation caused by FABP5 knockdown.
CONCLUSIONS: FABP5 promotes active fatty acid synthesis and excessive proliferation of GCs by activating PI3K-AKT signaling, suggesting that abnormally high expression of FABP5 in GCs may be a novel biomarker or a research target for PCOS treatment.
摘要：
背景：多囊卵巢综合征（PCOS）是育龄妇女中最复杂的内分泌疾病之一。颗粒细胞(GCs)的异常增殖是PCOS的重要缘由。本研究旨在探讨脂肪酸结合蛋白5(FABP5)在多囊卵巢综合征(PCOS)患者颗粒细胞(GC)增殖中的作用。
方法：FABP5基因，这与脂质代谢有关，通过对来自基因表达综合(GEO)数据库的GSE138518的基因表达谱的数据分析来鉴定。通过定量实时PCR（qRT-PCR）和蛋白质印迹法测量FABP5的表达水平。用细胞计数试剂盒-8(CCK-8)测定评价细胞增殖。Western印迹用于评估增殖标志物PCNA的表达，免疫荧光显微镜检测Ki67表达。此外，用尼罗红染色检测脂滴形成，qRT-PCR用于分析脂肪酸储存相关基因的表达。
结果：我们发现FABP5在PCOS患者和PCOS小鼠的卵巢GCs中上调。FABP5敲低抑制人颗粒样肿瘤细胞系（KGN）的脂滴形成和增殖，而FABP5过表达显着增强了脂滴形成和KGN细胞增殖。此外，我们确定FABP5敲低通过抑制AKT磷酸化抑制PI3K-AKT信号传导,FABP5过表达通过促进AKT磷酸化激活PI3K-AKT信号传导.最后,我们使用PI3K-AKT信号通路抑制剂LY294002,发现FABP5过表达对KGN细胞增殖和脂滴形成的促进作用受到抑制。相比之下,PI3K-AKT信号通路激动剂SC79显著挽救了FABP5敲低引起的KGN细胞增殖抑制和脂滴形成。
结论：FABP5通过激活PI3K-AKT信号促进活性脂肪酸合成和GC的过度增殖，提示FABP5在GCs中的异常高表达可能是PCOS治疗的新生物标志物或研究靶点。
方法：FABP5基因，这与脂质代谢有关，通过对来自基因表达综合(GEO)数据库的GSE138518的基因表达谱的数据分析来鉴定。通过定量实时PCR（qRT-PCR）和蛋白质印迹法测量FABP5的表达水平。用细胞计数试剂盒-8(CCK-8)测定评价细胞增殖。Western印迹用于评估增殖标志物PCNA的表达，免疫荧光显微镜检测Ki67表达。此外，用尼罗红染色检测脂滴形成，qRT-PCR用于分析脂肪酸储存相关基因的表达。
结果：我们发现FABP5在PCOS患者和PCOS小鼠的卵巢GCs中上调。FABP5敲低抑制人颗粒样肿瘤细胞系（KGN）的脂滴形成和增殖，而FABP5过表达显着增强了脂滴形成和KGN细胞增殖。此外，我们确定FABP5敲低通过抑制AKT磷酸化抑制PI3K-AKT信号传导,FABP5过表达通过促进AKT磷酸化激活PI3K-AKT信号传导.最后,我们使用PI3K-AKT信号通路抑制剂LY294002,发现FABP5过表达对KGN细胞增殖和脂滴形成的促进作用受到抑制。相比之下,PI3K-AKT信号通路激动剂SC79显著挽救了FABP5敲低引起的KGN细胞增殖抑制和脂滴形成。
结论：FABP5通过激活PI3K-AKT信号促进活性脂肪酸合成和GC的过度增殖，提示FABP5在GCs中的异常高表达可能是PCOS治疗的新生物标志物或研究靶点。
