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DNMT1甲基化调控导致的miR-30b-5p表达降低参与胃癌转移。

Decreased miR-30b-5p expression by DNMT1 methylation regulation involved in gastric cancer metastasis.

作者信息

Qiao Fengchang, Zhang Kun, Gong Pihai, Wang Ling, Hu Jiaojiao, Lu Sen, Fan Hong

机构信息

Key Laboratory of Developmental Genes and Human Diseases, Ministry of Education, Department of Genetics and Developmental Biology, The Medical School of Southeast University and the Institute of Life Science, Southeast University, Nanjing, 210009, China,

出版信息

Mol Biol Rep. 2014 Sep;41(9):5693-700. doi: 10.1007/s11033-014-3439-4. Epub 2014 Jun 10.

Abstract

miRNAs have emerged as crucial regulators in the regulation of development as well as human diseases, especially tumorigenesis. The aims of this study are to evaluate miR-30b-5p expression pattern and mechanism in gastric carcinogenesis due to which remains to be determined. Expression of miR-30b-5p was analyzed in 51 gastric cancer cases and 4 cell lines by qRT-PCR. The effect of DNA methylation on miR-30b-5p expression was assessed by MSP and BGS. In order to know whether DNMT1 increased miR-30b-5p promoter methylation, DNMT1 was depleted in cell lines AGS and BGC-823. The role of miR-30b-5p on cell migration was evaluated by wound healing assays. Decreased expression of miR-30b-5p was found in gastric cancer samples. In tumor, the expression level of miR-30b-5p was profound correlated with lymph node metastasis (P = 0.019). The level of miR-30b-5p may be restored by DNA demethylation and DNMT1 induced miR-30b-5p promoter methylation. In vitro functional assays implied that enforced miR-30b-5p expression affected cell migration, consistent with tissues analysis. Our findings uncovered that miR-30b-5p is significantly diminished in gastric cancer tissues, providing the first insight into the epigenetic mechanism of miR-30b-5p down-regulation, induced by DNMT1, and the role of miR-30b-5p in gastric cancer carcinogenesis. Overexpression of miR-30b-5p inhibited cell migration. Thus, miR-30b-5p may represent a potential therapeutic target for gastric cancer therapy.

摘要

微小RNA(miRNAs)已成为发育调控以及人类疾病尤其是肿瘤发生过程中的关键调节因子。本研究的目的是评估miR-30b-5p在胃癌发生中的表达模式及机制,其相关情况仍有待确定。通过qRT-PCR分析了51例胃癌病例和4种细胞系中miR-30b-5p的表达情况。采用甲基化特异性PCR(MSP)和全基因组甲基化测序(BGS)评估DNA甲基化对miR-30b-5p表达的影响。为了解DNA甲基转移酶1(DNMT1)是否增加miR-30b-5p启动子甲基化,在AGS和BGC-823细胞系中敲低DNMT1。通过伤口愈合实验评估miR-30b-5p对细胞迁移的作用。发现胃癌样本中miR-30b-5p表达降低。在肿瘤中,miR-30b-5p的表达水平与淋巴结转移显著相关(P = 0.019)。DNA去甲基化可恢复miR-30b-5p水平,且DNMT1可诱导miR-30b-5p启动子甲基化。体外功能实验表明,强制表达miR-30b-5p会影响细胞迁移,这与组织分析结果一致。我们的研究结果发现,miR-30b-5p在胃癌组织中显著降低,首次揭示了由DNMT1诱导的miR-30b-5p下调的表观遗传机制以及miR-30b-5p在胃癌发生中的作用。miR-30b-5p过表达可抑制细胞迁移。因此,miR-30b-5p可能是胃癌治疗的一个潜在靶点。

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