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侵袭性和非侵袭性无功能促性腺激素垂体瘤在LINE-1重复元件的DNA甲基化水平上存在差异。

Invasive and Noninvasive Nonfunctioning Gonadotroph Pituitary Tumors Differ in DNA Methylation Level of LINE-1 Repetitive Elements.

作者信息

Rusetska Natalia, Kober Paulina, Król Sylwia Katarzyna, Boresowicz Joanna, Maksymowicz Maria, Kunicki Jacek, Bonicki Wiesław, Bujko Mateusz

机构信息

Department of Molecular and Translational Oncology, Maria Sklodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland.

Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland.

出版信息

J Clin Med. 2021 Feb 3;10(4):560. doi: 10.3390/jcm10040560.

DOI:10.3390/jcm10040560
PMID:33546126
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7913198/
Abstract

PURPOSE

Epigenetic dysregulation plays a role in pituitary tumor pathogenesis. Some differences in DNA methylation were observed between invasive and noninvasive nonfunctioning gonadotroph tumors. This study sought to determine the role of DNA methylation changes in repetitive LINE-1 elements in nonfunctioning gonadotroph pituitary tumors.

METHODS

We investigated LINE-1 methylation levels in 80 tumors and normal pituitary glands with bisulfite-pyrosequencing. Expression of two LINE-1 open reading frames ( and ) was analyzed with qRT- in tumor samples and mouse gonadotroph pituitary cells treated with DNA methyltransferase inhibitor. Immunohistochemical staining against L1-ORF1p was also performed in normal pituitary glands and tumors.

RESULTS

Hypomethylation of LINE-1 was observed in pituitary tumors. Tumors characterized by invasive growth revealed lower LINE-1 methylation level than noninvasive ones. LINE-1 methylation correlated with overall DNA methylation assessed with HM450K arrays and negatively correlated with and expression. Treatment of αT3-1 gonadotroph cells with 5-Azacytidine clearly increased the level of and mRNA; however, its effect on LβT2 cells was less pronounced. Immunoreactivity against L1-ORF1p was higher in tumors than normal tissue. No difference in L1-ORF1p expression was observed in invasive and noninvasive tumors.

CONCLUSION

Hypomethylation of LINE-1 is related to invasive growth and influences transcriptional activity of transposable elements.

摘要

目的

表观遗传失调在垂体肿瘤发病机制中起作用。在侵袭性和非侵袭性无功能促性腺激素瘤之间观察到一些DNA甲基化差异。本研究旨在确定DNA甲基化变化在无功能促性腺激素垂体肿瘤中重复LINE-1元件中的作用。

方法

我们用亚硫酸氢盐焦磷酸测序法研究了80个肿瘤和正常垂体腺中的LINE-1甲基化水平。用qRT-分析了肿瘤样本和用DNA甲基转移酶抑制剂处理的小鼠促性腺激素垂体细胞中两个LINE-1开放阅读框（和）的表达。还在正常垂体腺和肿瘤中进行了针对L1-ORF1p的免疫组织化学染色。

结果

在垂体肿瘤中观察到LINE-1低甲基化。以侵袭性生长为特征的肿瘤显示LINE-1甲基化水平低于非侵袭性肿瘤。LINE-1甲基化与用HM450K阵列评估的总体DNA甲基化相关，与和表达呈负相关。用5-氮杂胞苷处理αT3-1促性腺激素细胞明显增加了和mRNA的水平；然而，其对LβT2细胞的作用不太明显。肿瘤中针对L1-ORF1p的免疫反应性高于正常组织。在侵袭性和非侵袭性肿瘤中未观察到L1-ORF1p表达的差异。

结论

LINE-1低甲基化与侵袭性生长有关，并影响转座元件的转录活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2eb9/7913198/516f18813e42/jcm-10-00560-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2eb9/7913198/863eb44bc109/jcm-10-00560-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2eb9/7913198/c1bd3808a520/jcm-10-00560-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2eb9/7913198/516f18813e42/jcm-10-00560-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2eb9/7913198/863eb44bc109/jcm-10-00560-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2eb9/7913198/c1bd3808a520/jcm-10-00560-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2eb9/7913198/516f18813e42/jcm-10-00560-g003.jpg

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