作者：吴炜，刘永亮，王彦，董晓柳，张利，马思伟，张欢，高华，董伟

单位：

1.北京市神经外科研究所中心实验室2.河北省唐山市人民医院神经外科3.河北省唐山市人民医院神经内科

Units：

1.Central Lab, Beijing Neurosurgical Institute, Beijing 100070, China 2.Department of Neurosurgery, Tangshan People's Hospital, Tangshan 063000, China 3.Department of Neurology, Tangshan People's Hospital, Tangshan 063000, China

关键词：

垂体肿瘤; 垂体腺瘤; Delta样蛋白1同源物; 母系表达基因;

Keywords：

Pituitary neoplasms; Pituitary adenoma; Delta like non-canonical Notch ligand 1; Maternally expressed gene

CLC：

DOI：

10.12114/j.issn.1008-5971.2023.00.152

Funds：

2022年度河北省中央引导地方科技发展资金项目（226Z7704G）

摘要：

目的 分析垂体腺瘤患者Delta样蛋白1同源物（DLK1）、母系表达基因（MEG）3、MEG8基因表达水平及其临床意义。方法 选取2015年6月至2019年12月于唐山市人民医院行手术切除的垂体腺瘤患者92例为研究对象。收集患者一般资料，采用RT-qPCR法检测肿瘤组织DLK1、MEG3、MEG8、POU类1同源框1(POU1F1)、类固醇生成因子1(SF-1)基因表达水平，采用免疫组织化学染色检测肿瘤组织生长激素（GH）、PIT1、SF-1表达情况，采用ELISA检测血清GH、胰岛素样生长因子1(IGF-1)水平。比较GH腺瘤与无功能腺瘤患者一般资料，比较不同临床特征垂体腺瘤患者肿瘤组织DLK1、MEG3、MEG8基因表达水平，分析垂体腺瘤患者肿瘤组织DLK1、MEG3、MEG8基因表达水平与肿瘤组织POU1F1、SF-1基因表达水平的相关性，比较不同肿瘤组织DLK1、MEG3、MEG8基因表达水平垂体腺瘤患者肿瘤组织GH、PIT1、SF-1表达情况，分析GH腺瘤患者肿瘤组织DLK1、MEG3、MEG8基因表达水平与血清GH、IGF-1水平的相关性。结果 GH腺瘤患者男性占比、年龄低于无功能腺瘤患者（P<0.05）。男性垂体腺瘤患者肿瘤组织DLK1、MEG8基因表达水平低于女性垂体腺瘤患者（P<0.05）。年龄≤52岁垂体腺瘤患者肿瘤组织DLK1基因表达水平高于年龄>52岁垂体腺瘤患者（P<0.05）。GH腺瘤患者肿瘤组织DLK1、MEG3、MEG8基因表达水平高于无功能腺瘤患者（P<0.05）。Pearson相关分析结果显示，垂体腺瘤患者肿瘤组织DLK1、MEG3、MEG8基因表达水平与肿瘤组织POU1F1基因表达水平均呈正相关，与肿瘤组织SF-1基因表达水平均呈负相关（P<0.05）。根据肿瘤组织DLK1、MEG3、MEG8基因表达水平的中位数，分别将患者分为DLK1基因高表达组（DLK1基因表达水平≥0.000 15,46例）和DLK1基因低表达组（DLK1基因表达水平<0.000 15,46例）、MEG3基因高表达组（MEG3基因表达水平≥0.000 11,46例）和MEG3基因低表达组（MEG3基因表达水平<0.000 11,46例）、MEG8基因高表达组（MEG8基因表达水平≥0.000 08,46例）和MEG8基因低表达组（MEG8基因表达水平<0.000 08,46例）。DLK1基因低表达组肿瘤组织GH、PIT1阳性者占比及SF-1弱阳性、阴性者占比低于DLK1基因高表达组，GH、PIT1阴性者占比及SF-1阳性者占比高于DLK1基因高表达组（P<0.05）;MEG3基因低表达组肿瘤组织GH、PIT1阳性者占比及SF-1弱阳性、阴性者占比低于MEG3基因高表达组，GH、PIT1阴性者占比及SF-1阳性者占比高于MEG3基因高表达组（P<0.05）;MEG8基因低表达组肿瘤组织GH、PIT1阳性者占比及SF-1弱阳性、阴性者占比低于MEG8基因高表达组，GH、PIT1阴性者占比及SF-1阳性者占比高于MEG8基因高表达组（P<0.05）。Pearson相关分析结果显示，GH腺瘤患者肿瘤组织DLK1基因表达水平与血清GH水平呈正相关（P<0.05），与血清IGF-1水平无直线相关关系（P>0.05）;GH腺瘤患者肿瘤组织MEG3、MEG8基因表达水平与血清GH、IGF-1水平均无直线相关关系（P>0.05）。结论 垂体腺瘤患者的DLK1、MEG3、MEG8基因表达水平低于正常人群。DLK1、MEG3、MEG8基因表达水平升高可促进垂体腺瘤细胞向PIT1谱系分化，临床表现为GH腺瘤表型；其降低可促进垂体腺瘤细胞向SF-1谱系分化，临床表现为无功能腺瘤表型。DLK1基因是调控GH分泌的关键基因，其可能作为GH腺瘤患者的治疗靶点。

Abstract：

Objective To analyze expression levels and clinical significance of Delta like non-canonical Notch ligand 1 (DLK1) , maternally expressed gene (MEG) 3 and MEG8 genes in patients with pituitary adenoma. Methods A total of 92 patients with pituitary adenoma who underwent surgical resection in Tangshan People's Hospital from June 2015 to December 2019 were selected as the research objects. General data of patients were collected, the expression levels of DLK1, MEG3, MEG8, POU class 1 homeobox 1 (POU1F1) and steroidogenic factor 1 (SF-1) genes in tumor tissues were detected by RT-qPCR. The expression of growth hormone (GH) , PIT1 and SF-1 in tumor tissues was detected by immunohistochemical staining. Serum GH and insulin like growth factor 1 (IGF-1) levels were determined by ELISA. The general data of patients with GH adenoma and non-functional adenoma were compared. The expression levels of DLK1, MEG3, and MEG8 genes in tumor tissues were compared in pituitary adenoma patients with different clinical characteristics. The correlation between the expression levels of DLK1, MEG3 and MEG8 genes and POU1F1 and SF-1 genes in tumor tissues of patients with pituitary adenoma was analyzed. The expression of GH, PIT1, and SF-1 in tumor tissues was compared in pituitary adenoma patients with different expression levels of DLK1, MEG3, MEG8 gene in tumor tissues. The correlation between the expression levels of DLK1, MEG3, MEG8 genes in tumor tissue and serum GH and IGF-1 levels in patients with GH adenoma was analyzed. Results The male proportion and age of GH adenoma patients were lower than those of non-functional adenoma patients (P < 0.05) . The expression levels of DLK1 and MEG8 genes in male pituitary adenoma patients were lower than those in female pituitary adenoma patients (P < 0.05) . The expression level of DLK1 gene in tumor tissues of pituitary adenoma patients aged ≤ 52 years was higher than that of pituitary adenoma patients aged > 52 years (P < 0.05) . The expression levels of DLK1, MEG3 and MEG8 genes in tumor tissues of GH adenoma patients were higher than those of non-functional adenoma patients (P < 0.05) . Pearson correlation analysis results showed that the expression levels of DLK1, MEG3, and MEG8 genes in tumor tissue of pituitary adenoma patients were positively correlated with the expression level of POU1F1 gene in tumor tissue, and negatively correlated with the expression level of SF-1 gene in tumor tissue (P < 0.05) . According to the median expression levels of DLK1, MEG3, and MEG8 genes in tumor tissue, patients were divided into DLK1 gene high expression group (DLK1 gene expression level ≥ 0.000 15, 46 cases) and DLK1 gene low expression group (DLK1 gene expression level < 0.000 15, 46 cases) , MEG3 gene high expression group (MEG3 gene expression level ≥ 0.000 11, 46 cases) and MEG3 gene low expression group (MEG3 gene expression level < 0.000 11, 46 cases) , MEG8 gene high expression group (MEG8 gene expression level ≥ 0.000 08, 46 cases) and MEG8 gene low expression group (MEG8 gene expression level < 0.000 08, 46 cases) . The proportions of GH and PIT1 positive and SF-1 weak positive and negative in tumor tissue of DLK1 gene low expression group were lower than those of DLK1 gene high expression group, while the proportions of GH and PIT1 negative and SF-1 positive were higher than those of DLK1 gene high expression group (P < 0.05) . The proportions of GH and PIT1 positive and SF-1 weak positive and negative in tumor tissue of MEG3 gene low expression group were lower than those of MEG3 gene high expression group, while the proportions of GH and PIT1 negative and SF-1 positive were higher than those of MEG3 gene high expression group (P < 0.05) . The proportions of GH and PIT1 positive and SF-1 weak positive and negative in tumor tissue of MEG8 low expression group were lower than those of MEG8 high expression group, while the proportions of GH and PIT1 negative and SF-1 positive were higher than those of MEG8 high expression group (P < 0.05) . Pearson correlation analysis showed that the expression level of DLK1 gene in tumor tissue of GH adenoma patients was positively correlated with serum GH level (P < 0.05) , but not linearly correlated with serum IGF-1 level (P > 0.05) . There was no linear correlation between the expression levels of MEG3 and MEG8 genes in tumor tissues and serum GH and IGF-1 levels in patients with GH adenoma (P > 0.05) . Conclusion The expression levels of DLK1, MEG3, and MEG8 genes in patients with pituitary adenoma are lower than those in the normal population. Elevated expression levels of DLK1, MEG3, and MEG8 genes can promote the differentiation of pituitary adenoma cells into the PIT1 lineage, clinically manifested as a GH adenoma phenotype; its reduction can promote the differentiation of pituitary adenoma cells into the SF-1 lineage, clinically manifested as a non functional adenoma phenotype. The DLK1 gene is a key gene that regulates GH secretion and may serve as a therapeutic target for GH adenoma patients.

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