APC gene is modulated by hsa-miR-135b-5p in both diffuse and intestinal gastric cancer subtypes.-猫眼课题宝

DOI：

10.1186/s12885-018-4980-7

发表时间：

2018-10-30

期刊：

BMC cancer

影响因子：

3.8

通讯作者：

中科院分区：

医学2区

文献类型：

作者：

Magalhães L;Quintana LG;Lopes DCF;Vidal AF;Pereira AL;D'Araujo Pinto LC;de Jesus Viana Pinheiro J;Khayat AS;Goulart LR;Burbano R;de Assumpção PP;Ribeiro-Dos-Santos Â

来源链接：

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208123/

一些遗传和表观遗传改变与胃癌（GC）的发生和进展相关，其中之一是去调节的microRNA（miRNA）表达谱。miRNA是一种小的非编码RNA，负调控成千上万的基因的表达，包括癌基因和肿瘤抑制基因。在先前的研究中，我们的小组鉴定了一些在GC中与无癌胃粘膜相比差异表达的miRNA，包括hsa-miR-29 c和hsa-miR-135 b。该研究的目的是调节miRNAs hsa-miR-29 c-5 p和hsa-miR-135 b-5 p的表达，并评估其靶基因在2D和3D细胞培养物中的表达。hsa-miR-29 c-5 p和hsa-miR-135 b-5 p的表达谱分别在2D和3D细胞培养物中通过转染模拟物和antimiR调节。Western Blot法检测hsa-miR-29 c-5 p靶基因CDC 42、DNMT 3A和hsa-miR-135 b-5 p靶基因APC蛋白的表达。结果显示，mimics和antimiRs转染显著改变了两种miRNAs的表达，增加了hsa-miR-29 c-5 p的表达，降低了hsa-miR-135 b-5 p的表达，尤其是在细胞系的三维培养中。当分析蛋白质表达时，我们观察到用模拟物转染的AGP 01和AGP 03细胞系具有CDC 42和DNMT 3A水平的降低，并且用antimiR转染的所有三种细胞系具有蛋白质APC表达的增加。我们的结论是，三维培养可以是一个更有代表性的体外模型，更好地模拟体内的现实。我们的研究结果还表明，hsa-miR-29 c-5 p是肠型胃癌中CDC 42和DNMT 3A基因的重要调节因子，而hsa-miR-135 b-5 p在肠型和弥漫型胃癌中均调节APC基因。它们的表达失调，从而在各自的信号通路，显示了这些miRNA如何影响不同组织学亚型的胃癌的发生。本文的在线版本（10.1186/s12885-018-4980-7）包含补充材料，可供授权用户使用。

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. 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 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. 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. The online version of this article (10.1186/s12885-018-4980-7) contains supplementary material, which is available to authorized users.