miR-101-2, miR-125b-2 and miR-451a act as potential tumor suppressors in gastric cancer through regulation of the PI3K/AKT/mTOR pathway.
miR-101-2, miR-125b-2 and miR-451a act as potential tumor suppressors in gastric cancer through regulation of the PI3K/AKT/mTOR pathway.
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DOI：
10.1007/s13402-015-0247-3
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发表时间：
2016-02
期刊：
影响因子：
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通讯作者：
Roa JC
中科院分区：
文献类型：
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作者：
Riquelme I;Tapia O;Leal P;Sandoval A;Varga MG;Letelier P;Buchegger K;Bizama C;Espinoza JA;Peek RM;Araya JC;Roa JC
胃癌是一种世界性的恶性肿瘤。在过去，它已被证明，细胞信号通路的改变在GC的发展中起着至关重要的作用。具体而言，PI 3 K/AKT/mTOR通路的失调似乎影响多种GC功能，包括生长、增殖、代谢、运动性和血管生成。通过microRNAs（miRNAs）靶向改变这一途径代表了一种潜在的治疗策略，特别是在耐药肿瘤中。本研究的目的是评估3种预先选择的miRNAs，miR-101-2，miR-125 b-2和miR-451 a在一系列原发性GC组织和匹配的非GC组织以及几种GC衍生的细胞系中的表达，并随后评估这些miRNAs的功能作用。25份原代GC样品、25份匹配的非GC样品和3份GC衍生细胞系，即，AGS、MKN 28和MKN 45包括在本研究中。分别通过定量RT-PCR和Western blotting评估miRNA和靶基因表达水平。随后，对AGS细胞进行细胞活力、克隆形成、细胞死亡、迁移和侵袭测定。发现miR-101-2、miR-125 b-2和miR-451 a在所测试的原代GC组织和GC衍生的细胞系中下调。在AGS细胞中，miRNA模拟物转染显著降低了细胞活力和集落形成，增加了细胞死亡并降低了细胞迁移和侵袭。我们还发现miR-101-2、miR-125 b-2和miR-451 a的外源性表达分别降低了其推定靶点MTOR、PIK 3CB和TSC 1的表达。我们的表达分析和体外功能测定表明，miR-101-2，miR-125 b-2和miR-451 a在原代GC以及GC衍生的AGS细胞中作为潜在的肿瘤抑制因子。
Gastric cancer (GC) is a deadly malignancy worldwide. In the past, it has been shown that cellular signaling pathway alterations play a crucial role in the development of GC. In particular, deregulation of the PI3K/AKT/mTOR pathway seems to affect multiple GC functions including growth, proliferation, metabolism, motility and angiogenesis. Targeting alterations in this pathway by microRNAs (miRNAs) represents a potential therapeutic strategy, especially in inhibitor-resistant tumors. The objective of this study was to evaluate the expression of 3 pre-selected miRNAs, miR-101-2, miR-125b-2 and miR-451a, in a series of primary GC tissues and matched non-GC tissues and in several GC-derived cell lines, and to subsequently evaluate the functional role of these miRNAs. Twenty-five primary GC samples, 25 matched non-GC samples and 3 GC-derived cell lines, i.e., AGS, MKN28 and MKN45, were included in this study. miRNA and target gene expression levels were assessed by quantitative RT-PCR and western blotting, respectively. Subsequently, cell viability, clone formation, cell death, migration and invasion assays were performed on AGS cells. miR-101-2, miR-125b-2 and miR-451a were found to be down-regulated in the primary GC tissues and the GC-derived cell lines tested. MiRNA mimic transfections significantly reduced cell viability and colony formation, increased cell death and reduced cell migration and invasion in AGS cells. We also found that exogenous expression of miR-101-2, miR-125b-2 and miR-451a decreased the expression of their putative targets MTOR, PIK3CB and TSC1, respectively. Our expression analyses and in vitro functional assays suggest that miR-101-2, miR-125b-2 and miR-451a act as potential tumor suppressors in primary GCs as well as in GC-derived AGS cells.