二甲双胍通过 miR-148/152 家族调控非小细胞肺癌细胞中 DNA 甲基转移酶的表达。
Metformin regulates expression of DNA methyltransferases through the miR-148/-152 family in non-small lung cancer cells.
机构信息
Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon, 16419, Korea.
Yonsei New I1 Han Institute for Integrative Lung Cancer Research, Yonsei University College of Medicine, Seoul, 03772, Korea.
出版信息
Clin Epigenetics. 2023 Mar 23;15(1):48. doi: 10.1186/s13148-023-01466-0.
BACKGROUND
To understand the molecular mechanisms involved in regulation of DNA methyltransferases (DNMTs) by metformin in non-small cell lung cancer (NSCLC) cells.
METHODS
Expression levels of DNMTs in response to metformin were analyzed in NSCLC cells. MicroRNAs regulating expression of DNMTs at the post-transcriptional level were searched using miRNA-target databases (miRDB and miRTarBase), TCGA RNASeqV2 lung cancer data, and miRNA-seq.
RESULTS
Metformin dose-dependently downregulated expression of DNMT1 and DNMT3a at the post-transcriptional level and expression of DNMT3b at the transcriptional level in A549 lung cancer cells. Activity of DNMTs was reduced by about 2.6-fold in A549 cells treated with 10 mM metformin for 72 h. miR-148/-152 family members (miR-148a, miR-148b, and miR-152) targeting the 3'UTR of DNMTs were associated with post-transcriptional regulation of DNMTs by metformin. Metformin upregulated expression of miR-148a, miR-148b, and miR-152 in A549 and H1650 cells. Transfection with an miR-148b plasmid or a mimic suppressed expression of DNMT1 and DNMT3b in A549 cells. Transfection with the miR-148a mimic in A549 and H1650 cells decreased the luciferase activity of DNMT1 3'UTR. A combination of metformin and cisplatin synergistically increased expression levels of miR-148/-152 family members but decreased expression of DNMTs in A549 cells. Low expression of miR-148b was associated with poor overall survival (HR = 2.56, 95% CI 1.09-6.47; P = 0.04) but not with recurrence-free survival.
CONCLUSIONS
The present study suggests that metformin inhibits expression of DNMTs by upregulating miR-148/-152 family members in NSCLC cells.
背景
为了了解二甲双胍在非小细胞肺癌（NSCLC）细胞中调节 DNA 甲基转移酶（DNMTs）的分子机制。
方法
分析 NSCLC 细胞中二甲双胍对 DNMTs 表达的影响。使用 miRNA 靶标数据库（miRDB 和 miRTarBase）、TCGA RNASeqV2 肺癌数据和 miRNA-seq 搜索在转录后水平调节 DNMTs 表达的 microRNAs。
结果
二甲双胍在 A549 肺癌细胞中呈剂量依赖性地下调 DNMT1 和 DNMT3a 的转录后表达，下调 DNMT3b 的转录表达。用 10 mM 二甲双胍处理 A549 细胞 72 h，DNMTs 的活性降低了约 2.6 倍。miR-148/-152 家族成员（miR-148a、miR-148b 和 miR-152）靶向 DNMTs 的 3'UTR，与二甲双胍对 DNMTs 的转录后调节有关。二甲双胍上调 A549 和 H1650 细胞中 miR-148a、miR-148b 和 miR-152 的表达。转染 miR-148b 质粒或 mimic 可抑制 A549 细胞中 DNMT1 和 DNMT3b 的表达。在 A549 和 H1650 细胞中转染 miR-148a 模拟物可降低 DNMT1 3'UTR 的荧光素酶活性。二甲双胍和顺铂联合使用可协同增加 miR-148/-152 家族成员的表达水平，但降低 A549 细胞中 DNMTs 的表达。miR-148b 低表达与总生存期较差相关（HR=2.56，95%CI 1.09-6.47；P=0.04），但与无复发生存期无关。
结论
本研究表明，二甲双胍通过上调 NSCLC 细胞中的 miR-148/-152 家族成员抑制 DNMTs 的表达。
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