Department of Urology, Shengli Clinical Medical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou, China.
Urol Int. 2023;107(5):517-525. doi: 10.1159/000522634. Epub 2022 Mar 30.
DNA methylation regulates gene transcriptional functions in the pathogenesis of malignant diseases. In prostate cancer, several tumor suppressors are known to be tumor specifically methylated.
In this study, 450K methylation data and mRNA expression data were accessed from The Cancer Genome Atlas-Prostate Adenocarcinoma database and analyzed bioinformatically. Methylation-specific PCR was used to examine the methylation condition in AOX1 promoter. qRT-PCR was applied to measure the mRNA expression of AOX1. Western blot was employed to detect the expressions of AOX1 and the EMT associated proteins. Transwell and scratch healing assays were used to examine the invasive and migratory abilities of the prostate cancer cells respectively.
AOX1 was lowly expressed and hypermethylated in the prostate cancer tissues and cells. Also, AOX1 was downregulated at protein level in prostate cancer cells. Knocking down AOX1 could promote cell migration and invasion in the prostate cancer cells. By using a DNA methylation inhibitor, 5-AzadC was found to promote the expression of AOX1 and reverse the promoting effects of short interfering RNA against AOX1 on cell migration and invasion.
This study suggested that DNA methylation and low AOX1 level might be biomarkers for prostate cancer.
DNA 甲基化调控恶性疾病发病机制中的基因转录功能。在前列腺癌中,已知几种肿瘤抑制因子被肿瘤特异性甲基化。
本研究从癌症基因组图谱-前列腺腺癌数据库中获取了 450K 甲基化数据和 mRNA 表达数据,并进行了生物信息学分析。采用甲基化特异性 PCR 检测 AOX1 启动子的甲基化状态。qRT-PCR 用于测量 AOX1 的 mRNA 表达。Western blot 用于检测 AOX1 和 EMT 相关蛋白的表达。Transwell 和划痕愈合实验分别用于检测前列腺癌细胞的侵袭和迁移能力。
在前列腺癌组织和细胞中,AOX1 表达水平低且呈高甲基化状态。此外,在前列腺癌细胞中 AOX1 的蛋白水平下调。敲低 AOX1 可促进前列腺癌细胞的迁移和侵袭。使用 DNA 甲基化抑制剂 5-AzadC 发现可促进 AOX1 的表达,并逆转针对 AOX1 的小干扰 RNA 对细胞迁移和侵袭的促进作用。
本研究表明 DNA 甲基化和 AOX1 水平降低可能是前列腺癌的生物标志物。