Low abundance of TFPI-2 by both promoter methylation and miR-27a-3p regulation is linked with poor clinical outcome in gastric cancer.

低丰度的 TFPI-2 启动子甲基化和 miR-27a-3p 调控与胃癌的不良临床预后有关。

  • 影响因子:1.49
  • DOI:10.1002/jgm.3166
  • 作者列表:"Geng G","Liu X","Xu A","Lu Z","Chen K","He J","Qi D","Yuan X
  • 发表时间:2020-01-26

BACKGROUND:The tumor suppressor role of tissue factor pathway inhibitor 2 (TFPI-2) has been reported in various tumors. This study aimed to improve the understanding of the oncogenic properties of TFPI-2 in gastric cancer. METHODS:Relative expression of TFPI-2 was determined by real-time polymerase chain reaction (PCR) and Western blot, respectively. Cell viability was measured with Cell Counting Kit-8 assay and proliferation was evaluated by colony formation assay. Cell apoptosis was assessed with caspase-3 activity kit and invasion was evaluated by transwell chamber assay. The methylation level of TFPI-2 promoter was assayed by methylation-specific PCR. The regulatory effect of miR-27a-3p on TFPI-2 was analyzed with luciferase reporter assay. The direct association between miR-27a-3p and TFPI-2 was shown by biotin-labelling pulldown. RESULTS:TFPI-2 was down-regulated in gastric cancer, which associated with unfavorable prognosis clinically. Ectopic introduction of TFPI-2 greatly compromised cell viability, colony formation, and invasive capacity and induced cell apoptosis simultaneously. The promoter region of TFPI-2 was extensively methylated in gastric cancer tissues in comparison with normal tissues, suggesting the epigenetic inhibition of TFPI-2 expression. We further identified that TFPI-2 functioned as sponge RNA against miR-27a-3p. Most importantly, miR-27a-3p-specific inhibitor significantly exerted tumor suppressor function akin to TFPI-2 itself, and the anti-tumoral activities were completely abolished by TFPI-2 knockdown. CONCLUSIONS:We discovered that the epigenetically suppressed TFPI-2 compromised sponging effects on miR-27a-3p in gastric cancer, which consequently and mechanistically contributed to tumor biology of gastric cancer.


背景: 组织因子途径抑制物 2 (TFPI-2) 的肿瘤抑制作用在多种肿瘤中已有报道。本研究旨在提高对胃癌 TFPI-2 致癌特性的认识。 方法: 采用实时荧光定量聚合酶链反应 (PCR) 和免疫印迹法 (Western blot) 检测 TFPI-2 的相对表达量。细胞活力用细胞计数试剂盒-8 法测定,增殖用集落形成法评价。用 caspase-3 活性试剂盒评估细胞凋亡,用 transwell 小室试验评估侵袭。用甲基化特异性 PCR 检测 TFPI-2 启动子甲基化水平。用荧光素酶报告基因技术分析 miR-27a-3p 对 TFPI-2 的调节作用。通过生物素标记下拉显示 miR-27a-3p 与 TFPI-2 之间的直接关联。 结果: TFPI-2 在胃癌中表达下调,临床预后不良。异位引入 TFPI-2 极大地损害了细胞活力、集落形成和侵袭能力,并同时诱导细胞凋亡。与正常组织相比,TFPI-2 的启动子区在胃癌组织中广泛甲基化,提示表观遗传抑制了 TFPI-2 的表达。我们进一步鉴定 TFPI-2 作为海绵 RNA 作用于 miR-27a-3p。最重要的是,miR-27a-3p-specific 抑制剂显著发挥肿瘤抑制功能,类似于 TFPI-2 本身,并且通过 TFPI-2 敲除完全消除了抗肿瘤活性。 结论: 我们发现表观遗传学抑制了 TFPI-2 对胃癌 miR-27a-3p 的海绵效应,从而在机制上促进了胃癌的肿瘤生物学。



作者列表:["Xie W","Chen C","Han Z","Huang J","Liu X","Chen H","Zhang T","Chen S","Chen C","Lu M","Shen X","Xue X"]

METHODS::Diffuse gastric cancer (DGC) is a lethal malignancy lacking effective systemic therapy. Among the most provocative recent results in DGC has been that the alter of the cellular cytoskeleton and intercellular adhesion. CD2-associated protein (CD2AP) is one of the critical proteins regulating cytoskeleton assembly and intercellular adhesion. However, no study has investigated the expression and biological significance of CD2AP in gastric cancer (GC) to date. Therefore, the aim of our study was to explore if the expression of CD2AP is associated with any clinical features of GC and to elucidate the underlying mechanism. Immunohistochemistry of 620 patient tissue samples indicated that the expression of CD2AP is downregulated in DGC. Moreover, a low CD2AP level was indicative of poor patient prognosis. In vitro, forced expression of CD2AP caused a significant decrease in the migration and invasion of GC cells, whereas depletion of CD2AP had the opposite effect. Immunofluorescence analysis indicated that CD2AP promoted cellular adhesion and influenced cell cytoskeleton assembly via interaction with the F-actin capping protein CAPZA1. Overall, the upregulation of CD2AP could attenuate GC metastasis, suggesting CD2AP as a novel biomarker for the prognosis and treatment of patients with GC.

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作者列表:["Jing JJ","Li H","Wang ZY","Zhou H","Sun LP","Yuan Y"]

METHODS::Aim: To identify the methylated-differentially expressed genes (MDEGs) that may serve as diagnostic markers and therapeutic targets in Epstein-Barr virus-associated gastric cancer (EBVaGC) and to explore the methylation-based pathways for elucidating biological mechanisms of EBVaGC. Materials & methods: Gene expression and methylation profiles were downloaded from GEO database. MDEGs were identified by GEO2R. Pathway enrichment analyses were conducted based on DAVID database. Hub genes were identified by Cytoscape, which were further verified by The Cancer Genome Atlas database. Results: A total of 367 hypermethylated, lowly expressed genes were enriched in specific patterns of cell differentiation. 31 hypomethylated, highly expressed genes demonstrated enrichment in regulation of immune system process. After validation using The Cancer Genome Atlas database, seven genes were confirmed to be significantly different hub genes in EBVaGC. Conclusion: EBVaGC-specific MDEGs and pathways can be served as potential biomarkers for precise diagnosis and treatment of EBVaGC and provide novel insights into the mechanisms involved.

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作者列表:["Daryabari SS","Fathi M","Mahdavi M","Moaddab Y","Hosseinpour Feizi MA","Shokoohi B","Safaralizadeh R"]

METHODS:Gastric adenocarcinoma, like other cancers, is a multifactorial genetic disease, andmetastasis of cancer cells is one of the main features of this illness. The expressionlevels of the CFL1 gene have been modulated in this pathway. Using small interferingRNA (siRNA) in the treatment of gastric cancer is considered a hopeful genetherapeutic approach. The present study reported the level of CFL1 genes betweentumor and margin and healthy tissue of gastric cancer. Also, the features of a cationicnanoparticle with a polymer coating containing polyacrylic acid and polyethylenei-mine that were used in the delivery of CFL1 siRNA, were shown. Then thecytotoxicity, cellular uptake, and gene silencing efficiency of this nanoparticle wereevaluated with CFL1siRNA. Method:In this study, the CFL1 gene expression was measured in 40 gastricadenocarcinoma, marginal and 15 healthy biopsy samples by a real‐time polymerasechain reaction. Physicochemical characteristics, apoptosis, and inhibition of migrationof the delivery of CFL1 siRNA by nanoparticle and lipofectamine were investigated ingastric cancer cells. Result:The CFL1 expression was remarkably increased in gastric cancer tissues incomparison with the marginal samples and normal tissues (p< .05) and the biomarkerindex for CFL1 was obtained as 0.94, then this gene can be probably used as abiomarker for gastric cancer. After treatment of the AGS cell line by CFL1 siRNA, theCFL1 expression level of mRNA and migration in AGS cells were remarkablysuppressed after transfection. Furthermore, the amount of apoptosis increased(p< .05). Conclusion:Our results demonstrated that CFL1 downregulation in AGS cells caninterdict cell migration. Finally, our outcomes propose that CFL1 can function as anoncogenic gene in gastric cancer and would be considered as a potential purpose ofgene therapy for gastric cancer treatment

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