APC 基因在弥漫型和肠型胃癌亚型中均受 hsa-miR-135b-5p 的调控。
APC gene is modulated by hsa-miR-135b-5p in both diffuse and intestinal gastric cancer subtypes.
机构信息
Laboratório de Genética Humana e Médica, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, Brazil.
Núcleo de Pesquisas em Oncologia, Hospital Universitário João de Barros Barreto, Universidade Federal do Pará, Belém, Brazil.
出版信息
BMC Cancer. 2018 Oct 30;18(1):1055. doi: 10.1186/s12885-018-4980-7.
BACKGROUND
Several genetic and epigenetic alterations are related to the development and progression of Gastric Cancer (GC), one of those being the deregulated microRNA (miRNA) expression profile. miRNAs are small noncoding RNAs that negatively regulate the expression of thousands of genes, including oncogenes and tumor suppressor genes. Our group identified, in previous studies, some miRNAs that are differentially expressed in GC when compared to the gastric mucosa without cancer, including hsa-miR-29c and hsa-miR-135b. The aim of the study was to modulate the expression of the miRNAs hsa-miR-29c-5p and hsa-miR-135b-5p and evaluate the expression of their target genes in 2D and 3D cell cultures.
METHODS
hsa-miR-29c-5p and hsa-miR-135b-5p expression profiles were modulated by transfecting mimics and antimiRs, respectively, in 2D and 3D cell cultures. The expression of the proteins coded by the genes CDC42, DNMT3A (target genes of hsa-miR-29c-5p) and APC (target gene of hsa-miR-135b-5p) were measured by Western Blot.
RESULTS
Results showed that mimics and antimiRs transfection significantly altered the expression of both miRNAs, increasing the expression of hsa-miR-29c-5p and reducing the expression of hsa-miR-135b-5p, especially in the 3D culture of the cell lines. When analyzing the proteins expression, we observed that AGP01 and AGP03 cell lines transfected with mimics had a reduction in the levels of CDC42 and DNMT3A and all three cell lines transfected with antimiRs had an increase in the expression of the protein APC.
CONCLUSION
We concluded that three-dimensional culture can be a more representative in vitro model that resembles better the in vivo reality. Our results also showed that hsa-miR-29c-5p is an important regulator of CDC42 and DNMT3A genes in the intestinal subtype gastric cancer and hsa-miR-135b-5p regulates the APC gene in both intestinal and diffuse subtypes of GC. Dysregulation in their expression, and consequently in their respectively signaling pathways, shows how these miRNAs can influence the carcinogenesis of different histological subtypes of gastric cancer.
背景
多种遗传和表观遗传改变与胃癌（GC）的发生和发展有关，其中之一是 miRNA 表达谱的失调。miRNA 是一种小的非编码 RNA，可负调控数千个基因的表达，包括癌基因和肿瘤抑制基因。我们的研究小组在之前的研究中发现，与无癌胃黏膜相比，GC 中存在一些差异表达的 miRNA，包括 hsa-miR-29c 和 hsa-miR-135b。本研究的目的是调节 miRNA hsa-miR-29c-5p 和 hsa-miR-135b-5p 的表达，并在 2D 和 3D 细胞培养物中评估其靶基因的表达。
方法
通过转染 mimic 和 antimiRs 分别调节 2D 和 3D 细胞培养物中 hsa-miR-29c-5p 和 hsa-miR-135b-5p 的表达谱。通过 Western Blot 测量由基因 CDC42、DNMT3A（hsa-miR-29c-5p 的靶基因）和 APC（hsa-miR-135b-5p 的靶基因）编码的蛋白质的表达。
结果
结果表明，miRNA mimics 和 antimiRs 的转染显著改变了两种 miRNA 的表达，增加了 hsa-miR-29c-5p 的表达，降低了 hsa-miR-135b-5p 的表达，尤其是在细胞系的 3D 培养物中。当分析蛋白质表达时，我们观察到转染 mimic 的 AGP01 和 AGP03 细胞系中 CDC42 和 DNMT3A 的水平降低，而转染 antimiRs 的所有三种细胞系中 APC 蛋白的表达增加。
结论
我们得出结论，三维培养可以是一种更具代表性的体外模型，更能模拟体内的真实情况。我们的结果还表明，hsa-miR-29c-5p 是肠型胃癌中 CDC42 和 DNMT3A 基因的重要调节因子，hsa-miR-135b-5p 调节 GC 肠型和弥漫型中 APC 基因。其表达失调及其相应的信号通路表明，这些 miRNA 如何影响不同组织学亚型胃癌的癌变。
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